Fast-Growing Alternative Food Materials: Current Status and Applications

Mealworms (Tenebrio molitor) are edible insects typically used as feed for captive reptiles, fish and birds. However, the incorporation of mealworm feed substrate waste (FSW) in fish feed has not been fully investigated. The objective of this study was to evaluate the nutritional composition and in vitro digestibility of mealworm FSW. Mealworm larvae were reared on a substrate of commercial chicken feed. Triplicate samples () of FSW were harvested on days 0 (0FSW), 7 (7FSW) and 14 (14FSW). The proximate chemical compositions of the three substrates were statistically different (). The FSW exhibited changes in proximate chemical composition, nutritional profile, and physicochemical properties that were related to the time of harvesting. Crude protein, crude fiber, and crude ash contents increased with time while ether extract, nitrogen-free extract, and gross energy decreased (). Fourier transform infrared spectrometry indicated some modifications in the nutritive profile of the FSW, and the pH, water solubility, water absorption capacity, microstructure, thermal characteristics and secondary protein structures changed. In vitro protein digestibility (IVPD) was highest in 7FSW in response to Nile tilapia (Oreochromis niloticus) digestive enzymes, and highest in 14FSW in response to striped catfish (Pangasianodon hypophthalmus) enzymes (). The IVPD values were generally lower than those of commercial fish feeds or commercial feed ingredients (). Our findings suggest that mealworm FSW could be included in aquafeed as an alternative protein source.

The house cricket (Acheta domesticus L.) is one of four edible insect species introduced to the EU market as a novel food and alternative protein source. Innovative products, such as cricket flour, are increasingly appearing on supermarket shelves and can offer an alternative to traditional cereals, while providing the body with many valuable nutrients of comparable quality to those found in meat and fish. The aim of this study was to investigate the possibility of using cricket powder as a substitute for wheat flour in the production of bread. The physicochemical properties of cricket powder were evaluated in comparison to wheat flour. As a result of technological studies, bread compositions with 5%, 10% and 15% replacements of wheat flour by cricket powder were designed and their quality characteristics (physicochemical, sensory and microbiological) were evaluated. Cricket powder was characterised by a higher protein (63% vs. 13.5%) and fat (16.3% vs. 1.16%) content and a lower carbohydrate (9.8% vs. 66%) and fibre (7.8% vs. 9.5%) content as compared to wheat flour. The tested preparations had a similar pH (6.9 and 6.8, respectively, for cricket powder and flour) and fat absorption capacity (0.14 vs. 0.27 g oil/g powder, respectively, for cricket powder and flour) but different water holding capacities and completely different colour parameters. All breads had good microbiological quality after baking and during 7 days of storage. In instrumental tests, the 10 and 15% replacements of wheat flour by cricket powder affected the darker colour of the breads and caused a significant increase in the hardness of the breads. The research has shown that the optimal level of replacement, which does not significantly affect the physiochemical and sensory characteristics, is 5% cricket powder in the bread recipe. Considering the results obtained and the fact that insects provide a sufficient supply of energy and protein in the human diet, are a source of fibre, vitamins and micronutrients, and have a high content of monounsaturated and polyunsaturated fatty acids, the suitability of cricket powder for protein enrichment of bakery products is confirmed.

The rapid growth of aquaculture has increased the demand for fish meal and fish oil, essential components of fish feed, leading to higher production costs due to dwindling global supplies. As the industry seeks sustainable alternatives, insects, particularly the yellow mealworm (Tenebrio molitor), are emerging as a viable protein source. With a nutritional profile comparable to fish meal and low environmental impact, insects offer an efficient and economical solution for feed production. The European Union's approval of mealworms for both animal feed and human consumption highlights their potential to support food security and environmental sustainability amidst rising global protein demands.

The increasing production of edible insects on an industrial scale makes it crucial to implement appropriate technologies after harvesting to process safe and high quality insect products. The aim of this work was to compare the impact of different drying treatments used in the production of flour from Tenebrio molitor larvae. The larvae were subjected to freeze-drying (FD), conventional drying (CD), microwave drying (MWD), microwave drying without freezing prior blanching (MWDL), and microwave drying with addition of 0.1% butylated hydroxytoluene (BHT) during the blanching of the larvae (MWDA). The studied parameters included water activity (aw), instrumental colour, chemical composition, lipid oxidative processes, antioxidant activity, as well as microbiological status. The freeze-drying and conventional drying of the larvae reduced the aw of the derived flours (p < 0.0001); however, their nutritional profile revealed lower protein (p < 0.0001) and considerably higher fat content (p < 0.0001) compared to the flours after microwave treatments. The conventional drying and microwave treatment with BHT induced significantly darker colour (p < 0.0001) in comparison to the other methods. Despite the advantages of the microwave drying as a fast and energy efficient method, it displayed some negative effects associated with low lipid stability such as higher acid value (AV) and secondary products of lipid oxidation (TBARS) (p < 0.0001). This was also observed in the MWDA flour, indicating a certain pro-oxidative effect of the BHT. Regardless of the drying method, all the flours had a low microbial load.

: Overcoming the planet’s food crisis is one of the greatest challenges today. The problem is even more acute when we think about the main foods consumed as a source of protein, such as meat, which is widely questioned because of the impacts its production generates. The world requires food that is nutritious, cheap

Abstract This comprehensive study investigated the diet’s impact on the growth and nutritional value of crickets (Acheta domesticus) in northern Thailand across five feed treatments. These included a commercial formula (CF21) with 21% protein content as a control, a novel cycling program of commercial formula (CF14) alternating between 21% and 14% protein feeds, and three new formulas (NF) with varied protein compositions (NF21 maintained a 21% protein diet, while NF17 and NF14 transitioned to 17% and 14% protein, respectively, after 21 days). Results revealed variations (p < 0.05) in production efficiency for 42-day-old crickets, including body weight, cricket production weight, and feed-to-body-weight conversion efficiency. NF21 displayed the highest average body weight for both sexes but the lowest total production weight. CF14 outperformed CF21 and the three new formulas. Despite trends in NF17 and NF14, their total weights slightly trailed CF14. Survival rates correlated with diet composition, with CF14 having the highest rate (71.08%), while subsequent groups experienced diminishing survival rates. Feed conversion ratio (FCR) analysis showed disparities, with NF21 recording the highest FCR value (3.34 ± 0.02) and CF14 recording the lowest (2.35 ± 0.01), highlighting different diet efficiencies. Economically, NF14 demonstrated cost-effectiveness at 25.92 ± 0.15 THB/kg (1 THB = 0.025 EUR), while CF14 emerged as the leader, achieving the highest net profit with a 51.51% cost reduction. The nutrient composition study revealed no significant impact on six values across all treatments. Overall, the majority of results favored CF14 as the preferred choice for achieving both cost-effectiveness and maximum yield.

Cordycepin production in the submerged culture of Cordyceps militaris was demonstrated using hydrolyzed corn processing protein by-products, known as corn steep liquor hydrolysate (CSLH), as an alternative nitrogen source. The growth, metabolism, and cordycepin production of Cordyceps militaris were evaluated under various concentrations of CSLH induction. The results demonstrated that CSLH addition had positive effects on the growth and cordycepin production with various C. militaris strains. The optimum strain, C. militaris GDMCC5.270, was found to effectively utilize CSLH to promote mycelium growth and cordycepin production. Low concentrations of CSLH (1.5 g/L) in the fermentation broth resulted in 343.03 ± 15.94 mg/L cordycepin production, which was 4.83 times higher than that of the group without CSLH. This also enhanced the metabolism of sugar, amino acids, and nucleotides, leading to improved cordycepin biosynthesis. The increase in key amino acids, such as glutamic acid, alanine, and aspartic acid, in the corn steep liquor hydrolysate significantly enhanced cordycepin yield. The corn steep liquor hydrolysate was confirmed to be a cost-effective accelerator for mycelium growth and cordycepin accumulation in C. militaris, replacing partial peptone as a cheap nitrogen source. It serves as a suitable alternative for efficient cordycepin production at a low cost.

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Among the different agricultural activities, the livestock is one of the most impacting on the environment. The feeding of animals is often the main responsible of the adverse environmental impact related to animal productions. Above all for intensive production, the consumption of protein feed is a key aspect for the achievement of sustainable production processes. The actual consumption of soybean meal and fish meal is not sustainable due to the related environmental impact and to the increasing prices. Among the different alternative protein sources, in the last 20 years, the attention of research centres and private companies focused on insects, algae and other invertebrates but, up to now, little consideration was paid to the use of fresh earthworm or earthworm meal as a protein feed for monogastric animals. The use of earthworms as an alternative protein source for fish and poultry feeding is an opportunity for providing environmental services via cleaner technologies. Thanks to earthworms, organic wastes and by-products generated by livestock activities can be valorised and become a resource for animal feeding in a circular perspective. In this context, this manuscript was designed to summarize the productivity, suitability and effectiveness issues connected with the utilization of earthworms as alternative protein feed in poultry production as well as in aquaculture. The studies investigating the earthworm meal use are quite old above all those carried out in Europe; however, some general indications can be drawn: both for broiler and fish, the parameters usually evaluated are body weight gain, growth rate, feed intake and feed conversion rate, the acceptability level of earthworm meal in broiler diet is lower than 15% while in trout diet ranges between 25 and 30%. The inclusion of earthworm meal in diets with an inclusion level lower than the acceptability threshold allows good productive performances without affecting the quality of the final food products.

Food production will have to increase significantly to meet the nutritional needs of the global population. There is also an urgent need to increase the sustainability of food production. Microalgae are a potential sustainable alternative to conventional protein sources and they can also be used in other industries such as agriculture or aquaculture. In this work, the cyanobacterium Arthrospira platensis was produced in Almeria (Spain) in a pilot-scale reactor (80 m2). The biomass produced was used as a protein source and a plant biostimulant following a biorefinery approach. Biomass productivity reached 5.6 g m−2 day−1. The biomass was rich in proteins (67.8 g (100 g)−1) and pigments, namely chlorophyll (7.6 mg (100 g)−1) and phycocyanin (134.2 mg (100 g)−1). An isoelectric solubilisation/precipitation method assisted by ultrasound led to the recovery of a protein extract with a protein content of 91.3 g (100 g)−1. The protein isolate was evaluated as a source of essential amino acids in tagliatelle, leading to an increase in the content of histidine, leucine, lysine, methionine, phenylalanine, threonine, and valine of 36.3, 75.2, 26.3, 30.0, 45.7, 57.8, and 70.0%, respectively. The protein content also increased from 9.6 to 13.9 g (100 g)−1 when the protein isolate was incorporated at a flour substitution level of 4%. The leftovers from the protein extraction were evaluated as plant biostimulants, for which auxin- and cytokinin-like effects were observed. Root development was especially promoted. The results demonstrated the feasibility of producing Spirulina during the winter in Europe and the potential simultaneous use of the biomass as a food ingredient and as a plant biostimulant.

The human population is growing and, globally, we must meet the challenge of increased protein needs required to feed this population. Single cell proteins (SCP), when coupled to aquaculture production, offer a means to ensure future protein needs can be met without direct competition with food for people. To demonstrate a given type of SCP has potential as a protein source for use in aquaculture feed, a number of steps need to be validated including demonstrating that the SCP is accepted by the species in question, leads to equivalent survival and growth, does not result in illness or other maladies, is palatable to the consumer, is cost effective to produce and can easily be incorporated into diets using existing technology. Here we examine white shrimp (Litopenaeus vannamei) growth and consumer taste preference, smallmouth grunt (Haemulon chrysargyreum) growth, survival, health and gut microbiota, and Atlantic salmon (Salmo salar) digestibility when fed diets that substitute the bacterium Methylobacterium extorquens at a level of 30% (grunts), 100% (shrimp), or 55% (salmon) of the fishmeal in a compound feed. In each of these tests, animals performed equivalently when fed diets containing M. extorquens as when fed a standard aquaculture diet. This transdisciplinary approach is a first validation of this bacterium as a potential SCP protein substitute in aquafeeds. Given the ease to produce this SCP through an aerobic fermentation process, the broad applicability for use in aquaculture indicates the promise of M. extorquens in leading toward greater food security in the future.

Due to the climate change crisis, and environmental impacts of the traditional meat sector, the production of artificial animal protein based on in vitro cell culture technology is proposed as an alternative. Furthermore, since traditional animal serum-supplemented cultures pose scientific challenges such as batch variation and contamination risks, artificial animal protein cultures are currently in urgent need of not only serum-free cultures, but also microcarrier culture systems for scalability. However, serum-free microcarrier-based culture system for the differentiation of muscle cells is not available to date. Therefore, we established an edible alginate microcapsules culture system for the differentiation of C2C12 cells in serum-free conditions. Furthermore, metabolites related to central carbon metabolism were profiled based on targeted metabolomics using mass spectrometry. The C2C12 cells cultured in alginate microcapsules displayed high viability throughout 7 days and successfully differentiated within 4 days in serum and serum-free cultures except for AIM-V cultures, which was confirmed by CK activity and MHC immunostaining. Lastly, to the best of our knowledge, this is the first report to compare metabolite profiles between monolayer and alginate microcapsule culture systems. Alginate microcapsule culture showed higher levels of intracellular glycolysis and TCA cycle intermediates, lactate, and the contribution of essential amino acids compared to the monolayer culture. We believe our serum-free alginate microcapsule culture system is adaptable to different species of muscle cells and contributes to future food technology as a proof of concept for the scalability of alternative animal protein source production.

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Duckweed has gained significant attention as a sustainable protein source due to its rapid growth, high protein content (up to 45 % dw), and well-balanced amino acid profile. Its potential as alternative to traditional plant-based proteins is explored, focusing on the extraction and purification methods that enhance protein yield and bioactivity. Techniques, including ultrasound-assisted extraction, high-pressure processing, and pulsed electric field, are highlighted for their ability to improve protein efficiency and functionality. Duckweed proteins exhibit valuable properties such as emulsification, foaming, and gelling, along with potential to produce bioactive peptides with health-promoting benefits. However, challenges such as heavy metal contamination, allergenicity, and the need for pure production remain barriers to large-scale commercialisation. A systematic search was carried to write this review article using PRISMA. Future research is essential to develop bioengineering solutions and optimise cultivation methods to fully explore duckweed's potential in contributing to sustainable food systems and the circular economy.

Alternative protein sources are gaining attraction in food industry and consumers. Proteins obtained by single-cell organisms, such as torula yeast, are of enormous interest, as they are highly scalable, efficient, and sustainable, and the production costs are comparably low. Nevertheless, proteins obtained from yeasts are still mostly known and studied for feed applications, despite their nutritional, functional, and sensory benefits for various food applications. Testing consumer acceptance of products, especially products containing alternative proteins provides insights into, e.g., market success, consumer perception, and optimization potential. In this study, the development of two vegan spread powders, high in protein and containing torula yeast as an alternative protein source, is introduced. The result of food product development using torula yeast were “Leberwurst”-style (14.7% protein) and a “Balkan”-style (9.7% protein) spreads both meeting the criteria “at least 20% kcal from proteins of total product kcal” and thus claimable as “high-protein.” The application of the alternative protein from torula yeast within the final products was studied by a consumer acceptance test (n = 123) within three different countries (Germany, Iceland, and Sweden). Consumers also rated their trust in food production actors, the food industry in particular, and their willingness to try new foods. Overall, both spreads received acceptance values in the range of “like slightly.” It is noticeable that the consumers liked the spread “Balkan style” more than “Leberwurst”-style. The background variables revealed higher neophobic characteristics of Icelandic consumers compared with Swedish or German consumers. However, German consumers felt transparency, and communication was missing, but Icelandic consumers, in general, had more trust in the overall food value chain. This knowledge allows for the development of strategies that address cultural-specific barriers and capitalize on cultural values that promote openness to culinary innovation. The identification of cultural variations in consumer preferences emphasizes the need for customized approaches to product development and marketing. These findings could have implications for businesses and policymakers in understanding and catering to the preferences and concerns of consumers in these respective countries. Businesses might benefit from emphasizing transparency and improving communication strategies. This could involve providing clear information about the sourcing, production, and other aspects of the food value chain.

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Cultivated meat (CM) technology has the potential to disrupt the food industry—indeed, it is already an inevitable reality. This new technology is an alternative to solve the environmental, health and ethical issues associated with the demand for meat products. The global market longs for biotechnological improvements for the CM production chain. CM, also known as cultured, cell-based, lab-grown, in vitro or clean meat, is obtained through cellular agriculture, which is based on applying tissue engineering principles. In practice, it is first necessary to choose the best cell source and type, and then to furnish the necessary nutrients, growth factors and signalling molecules via cultivation media. This procedure occurs in a controlled environment that provides the surfaces necessary for anchor-dependent cells and offers microcarriers and scaffolds that favour the three-dimensional (3D) organisation of multiple cell types. In this review, we discuss relevant information to CM production, including the cultivation process, cell sources, medium requirements, the main obstacles to CM production (consumer acceptance, scalability, safety and reproducibility), the technological aspects of 3D models (biomaterials, microcarriers and scaffolds) and assembly methods (cell layering, spinning and 3D bioprinting). We also provide an outlook on the global CM market. Our review brings a broad overview of the CM field, providing an update for everyone interested in the topic, which is especially important because CM is a multidisciplinary technology.

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As the global population grows, sustainable food production is crucial. Traditional livestock farming drives environmental degradation, while cultivated meat offers a promising alternative through cellular agriculture. Advances in tissue engineering and 3D bioprinting accelerate development, but challenges remain, including regulatory approval, scalability, intellectual property, and consumer acceptance. Ethical and social concerns, such as impacts on farmers and the use of animal‐derived media, further complicate commercialization. Collaboration among academia, industry, and policymakers is essential to overcoming these barriers. This perspective highlights key challenges and opportunities, emphasizing the need for strategic investment and regulation to realize cultivated meat's potential.

The growing world population, public awareness of animal welfare, environmental impacts and changes in meat consumption leads to the search for novel approaches to food production. Novel foods include products with a new or specifically modified molecular structure, foods made from microorganisms, fungi, algae or insects, as well as from animal cell or tissue cultures. The latter approach is known by various names: “clean meat”, “in vitro meat” and “cell‐cultured” or “(cell‐)cultivated meat”. Here, cells isolated from agronomically important species are expanded ex vivo to produce cell biomass used in unstructured meat or to grow and differentiate cells on scaffolds to produce structured meat analogues. Despite the fast‐growing field and high financial interest from investors and governments, cultivated meat production still faces challenges ranging from cell source choice, affordable expansion, use of cruelty‐free and food‐grade media, regulatory issues and consumer acceptance. This overview discusses the above challenges and possible solutions and strategies in the production of cultivated meat. The review integrates multifaceted historical, social, and technological insights of the field, and provides both an engaging comprehensive introduction for general interested and a robust perspective for experts.

Cellular agriculture is an emerging scientific discipline that leverages the existing principles behind stem cell biology, tissue engineering, and animal sciences to create agricultural products from cells in vitro. Cultivated meat, also known as clean meat or cultured meat, is a prominent subfield of cellular agriculture that possesses promising potential to alleviate the negative externalities associated with conventional meat production by producing meat in vitro instead of from slaughter. A core consideration when producing cultivated meat is cell sourcing. Specifically, developing livestock cell sources that possess the necessary proliferative capacity and differentiation potential for cultivated meat production is a key technical component that must be optimized to enable scale-up for commercial production of cultivated meat. There are several possible approaches to develop cell sources for cultivated meat production, each possessing certain advantages and disadvantages. This review will discuss the current cell sources used for cultivated meat production and remaining challenges that need to be overcome to achieve scale-up of cultivated meat for commercial production. We will also discuss cell-focused considerations in other components of the cultivated meat production workflow, namely, culture medium composition, bioreactor expansion, and biomaterial tissue scaffolding.

Cultured meat (also referred to as cultivated meat or cell-based meat)—CM—is fabricated through the process of cellular agriculture (CA), which entails application of bioengineering, i.e., tissue engineering (TE) principles to the production of food. The main TE principles include usage of cells, grown in a controlled environment provided by bioreactors and cultivation media supplemented with growth factors and other needed nutrients and signaling molecules, and seeded onto the immobilization elements—microcarriers and scaffolds that provide the adhesion surfaces necessary for anchor-dependent cells and offer 3D organization for multiple cell types. Theoretically, many solutions from regenerative medicine and biomedical engineering can be applied in CM-TE, i.e., CA. However, in practice, there are a number of specificities regarding fabrication of a CM product that needs to fulfill not only the majority of functional criteria of muscle and fat TE, but also has to possess the sensory and nutritional qualities of a traditional food component, i.e., the meat it aims to replace. This is the reason that bioengineering aimed at CM production needs to be regarded as a specific scientific discipline of a multidisciplinary nature, integrating principles from biomedical engineering as well as from food manufacturing, design and development, i.e., food engineering. An important requirement is also the need to use as little as possible of animal-derived components in the whole CM bioprocess. In this review, we aim to present the current knowledge on different bioengineering aspects, pertinent to different current scientific disciplines but all relevant for CM engineering, relevant for muscle TE, including different cell sources, bioreactor types, media requirements, bioprocess monitoring and kinetics and their modifications for use in CA, all in view of their potential for efficient CM bioprocess scale-up. We believe such a review will offer a good overview of different bioengineering strategies for CM production and will be useful to a range of interested stakeholders, from students just entering the CA field to experienced researchers looking for the latest innovations in the field.

The growing global demand for sustainable and safe food is a major challenge that increases the need for advanced alternatives such as tissue engineering (TE). TE offers promising solutions by improving yields, nutritional value and resilience of crops while also producing cultivated meat that reduces the environmental impact of livestock farming. The market potential for TE in meat production is considerable, and significant growth is expected. However, the regulatory framework for these innovations is developing slowly, and approval procedures vary across regions. This overview critically assesses the current applications of TE in the agri-food sector, their socio-economic potential and the regulatory challenges. It emphasises the need for harmonised, flexible and adaptive policies to effectively integrate engineered foods into the market.

Cellular agriculture combines cell culture and tissue engineering techniques to produce animal-derived food products. While cell culture is routinely reported in the medical literature to affect protein expression in primary mammalian cells, the impact of these changes has yet to be established in the context of food production and allergenicity. To address this, primary bovine myoblasts were cultured for short-, medium-, and long-durations and then compared to native tissue. Proteomic and immunoblotting analyses identified cumulative changes at increasing passage numbers, with significant differences between cultured cells and native tissue. While allergens Bos d 6 and Bos d 13 were less abundant in cultured cells, an increase in immunoglobulin E interactions with Galα1-3Galβ1-4GlcNAc-R (α-Gal) in cultured cells indicates that α-Gal may be more abundant. Results demonstrate that the relative abundance of allergens in cultivated beef products will likely differ from that of traditional meat, highlighting a need for further investigation.

To engineer full-thickness cultivated meat, a perfusable scaffold is required with a high density of capillaries that allow diffusion of oxygen, nutrients, and waste. Nie et al., established a remarkable feat of tissue engineering that provides important direction towards full-thickness cultivated meat.

Cultivated meat is a fast-growing research field and an industry with great potential to overcome the limitations of traditional meat production. Cultivated meat utilizes cell culture and tissue engineering technologies to culture a vast number of cells in vitro and grow/assemble them into structures mimicking the muscle tissues of livestock animals. Stem cells with self-renewal and lineage-specific differentiation abilities have been considered one of the key cell sources for cultivated meats. However, the extensive in vitro culturing/expansion of stem cells results in a reduction in their abilities to proliferate and differentiate. Extracellular matrix (ECM) has been used as a culturing substrate to support cell expansion for cell-based therapies in regenerative medicine due to its resemblance to the native microenvironment of cells. In this study, the effect of the ECM on the expansion of bovine umbilical cord stromal cells (BUSC) in vitro was evaluated and characterized. BUSCs with multi-lineage differentiation potentials were isolated from bovine placental tissue. Decellularized ECM prepared from a confluent monolayer of bovine fibroblasts (BF) is free of cellular components but contains major ECM proteins such as fibronectin and type I collagen and ECM-associated growth factors. Expansion of BUSC on ECM for three passages (around three weeks) resulted in about 500-fold amplification, while cells were amplified less than 10-fold when cultured on standard tissue culture plates (TCP). Moreover, the presence of ECM reduced the requirement for serum in the culture medium. Importantly, the cells amplified on ECM retained their differentiation abilities better than cells cultured on TCP. The results of our study support the notion that monolayer cell-derived ECM may be a strategy to expand bovine cells in vitro effectively and efficiently.

Cultivated meat offers a sustainable alternative to conventional livestock production, yet caprine species remain underexplored. This study investigates the bipotential differentiation capacity of muscle satellite cells (MuSCs) isolated from Aleppo goats (Capra hircus), aiming to optimize dual myogenic-adipogenic tissue generation from a single cell source. MuSCs were isolated via mechanical and enzymatic digestion, displaying spindle-shaped morphology and positive expression of CD56, while negative for CD31. To investigate dual-lineage specification, five co-culture models were established, two simultaneous (M1: mixed co-culture, M2: layered co-culture) and three sequential (M3: adipogenic-to-myogenic, M4: same as M3 but long adipogenic priming, M5: myogenic-to-adipogenic). Simultaneous models involved combining or layering lineage-committed cells, while sequential models exposed a single MuSC population to staged adipogenic and myogenic cues. Gene expression analysis and histological staining revealed that M3 yielded optimal myogenic differentiation, characterized by high MyoD and MyoG expression. Model 5 favored adipogenesis, as evidenced by PPARγ expression and lipid accumulation alongside late myogenic traits. These findings present an efficient strategy for generating tailored muscle-fat constructs from caprine MuSCs and highlight their potential for cost-effective, sustainable cultivated meat production.

Summary Cultivated meat has the potential to revolutionize food production, but its progress is hindered by fundamental shortcomings of mammalian cells with respect to industrial-scale bioprocesses. Here, we discuss the essential role of cell line engineering in overcoming these limitations, highlighting the balance between the benefits of enhanced cellular traits and the associated regulatory and consumer acceptance challenges. We believe that careful selection of cell engineering strategies, including both genetic and non-genetic modifications, can address this trade-off and is essential to advancing the field.

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The escalating environmental impact of traditional livestock farming, particularly beef production, has spurred the search for sustainable meat alternatives. This study introduces a novel Plant-Based Tissue Engineering (PBTE) approach, to replicate the complex structure and sensory experience of whole-muscle cuts of meat using plant-based ingredients. Leveraging principles of tissue engineering and advanced food manufacturing technologies, PBTE deconstructs meat into its fundamental components: muscle, fat, and connective tissue, and reconstructs them using a combination of plant proteins, fats and polysaccharide materials. The muscle component is reassembled to mimic the anisotropic fibrous structure of beef, while the fat component is engineered through lipid encapsulation within a hydrocolloid matrix. Advanced manufacturing techniques, including additive manufacturing and robotics, are utilized for precise spatial configuration and assembly of these components. Our findings demonstrate that PBTE can effectively replicate the mechanical integrity, texture, and sensory attributes of traditional meat, presenting a promising alternative that could significantly reduce the environmental footprint of meat production. This approach aligns with the principles of Soft Matter in the manipulation of artificial structures and materials for mimicking naturally occurring designs, such as whole cut meat foods. It also holds substantial potential for revolutionizing the alternative protein industry by catering to a broader consumer base, including flexitarians and meat-eaters.

The emerging field of cellular agriculture has accelerated the development of cell-cultivated adipose tissue as an additive to enhance the flavor of alternative meat products. However, there has been limited research to evaluate the sensory profile of in vitro-grown tissues compared to conventionally obtained animal fat. This study aimed to investigate the aromatic characteristics of cell-cultivated fat tissue as a flavor enhancer for meat alternatives. Porcine dedifferentiated fat cells were clonally isolated and differentiated into adipocytes. This cultured adipose tissue was then analyzed alongside native porcine fat using gas chromatography-mass spectrometry (GC/MS) coupled with descriptive sensory analysis by human panelists. This evaluation enabled quantitative and qualitative assessments of volatile compounds released during cooking for both in vitro and in vivo porcine fats. The volatile profiles generated during the cooking process and fatty aroma characteristics reported by sensory panelists were largely similar between the two fat sources, with some differences in the concentration of select compounds and aroma attributes. Ultimately, the panelists found comparable overall liking scores reported between the conventional and cultured porcine fats. These findings provide valuable sensory evidence supporting the viability of cell-cultivated adipose tissue as a flavor component of meat alternatives, substituting for conventional animal fat.

Livestock farming and conventional meat production pose significant environmental, health, and animal welfare challenges. In seeking sustainable alternative solutions, cultivated meat technology typically utilizes differentiation of myogenic progenitor cells (MPCs) into muscle cells for in vitro meat production. However, understanding the molecular determinants governing MPC differentiation into muscle cells, and the potential enhancement of this process through modulation of signaling pathways, remains limited. Herein, we characterized the molecular landscape associated with bovine MPC differentiation in vitro by employing multiomics, and explored its augmentation by small molecules, together leading to identification of media that enhanced myogenic differentiation compared with conventional methods in both 2D cultures and tissue‐engineered 3D skeletal muscle constructs. Through bulk and single‐cell transcriptomics and proteomics, we compared conventional and enhanced differentiation media, demonstrating that the enhanced media gave rise to unique progenitor‐like cell populations, while simultaneously promoting differentiation into myocytes and contractile myotubes expressing a wide array of myogenic markers that more closely resemble bovine muscle cells in vivo. The improved method for promoting myogenic differentiation in 2D and 3D formats, together with the corresponding molecular roadmap, may prove valuable for cultivated meat applications.

Cultured meat aims to replicate the sensory and functional properties of conventional meat by developing structured muscle tissue using cell culture. This study focuses on the culture of chicken embryonic and mesenchymal stem cells (MSCs) to derive muscle, and fat, optimizing conditions for differentiation and integration. We utilized monolayer and three-dimensional microcarrier-based cultures to produce muscle fibers and adipocytes while maintaining the extracellular matrix (ECM) integrity essential for tissue cohesion. Key pluripotency and myogenic markers (e.g., cOCT4, cMYOD, cMYH1E) were analyzed during differentiation, revealing dynamic gene expression patterns that underscore myogenesis. Myoblast differentiation into mature myotubes demonstrated decreased cPAX7 and increased cMYMK, confirming lineage commitment and muscle fiber formation. Adipogenesis was induced in MSCs using food-grade lecithin, which activated PPARγ, C/EBPα, and FABP4, resulting in robust lipid droplet accumulation. To scale production, microcarriers facilitated cell proliferation, while transglutaminase-based stabilization enabled the formation of three-dimensional tissue structures comparable to native meat. Our findings highlight advances in culture protocols, genotypic and phenotypic expression analyses of multinucleated chicken muscle and adipocyte cells for cultured meat production.

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Cultivating fat for edible tissue presents significant challenges, due to the high costs associated with growth and differentiation factors, alongside the poor viability of adipocytes resulting from cell clustering. Additionally, there is a gap in research regarding the rapid accumulation of fats within cells. To that end, this study presents the development of a biodegradable soy protein colloidosome system for an efficient application: direct delivery of oils into bovine satellite cells, enabling rapid intracellular fat accumulation without the need for adipogenic differentiation. This method aims to enhance fat content while maintaining high cell viability. Colloidosomes are prepared from paraffin/olive oil-in-water emulsions stabilized by soy protein nanoparticles. Their size, shape, and zeta potential are characterized by SEM, Cryo-TEM, and DLS analyses. Confocal microscopy, Cryo-SEM, and analytical centrifugation reveal the morphology, structure, and stability of the colloidosome shell structure. Bioassays on satellite cells show a 20-fold increase in AdipoRed signal when treated with colloidosomes containing 30 μL of olive oil, along with an 80% survival rate after 3 days compared to controls. Through the uptake of colloidosomes, cells act as "oil-rich cells," serving as an alternative form of fat. This method offers a fully biocompatible, safe, efficient, and cost-effective approach to the production of cultured fat, demonstrating a promising strategy for scaling up cultivated meat.

Cultivated meat (CM) has transitioned from a futuristic concept to a present reality, with select products approved for consumption and sale in Singapore, Israel, and the USA. This evolution has emphasized scalable, cost-effective, and sustainable production, as well as navigation of regulatory pathways. As CM develops, a crucial challenge lies in delivering products that are highly appealing to consumers. Central to this will be refining CM palatability, a term encompassing food's taste, aroma, texture, tenderness, juiciness, and color. We explore the scientific and engineering approaches to producing palatable CM, including cell-line selection, cell differentiation, and post-processing techniques. This includes a discussion of the structural and compositional properties of meat that are intrinsically coupled to palatability.

Alternative technology for meat production holds the potential to alleviate ethical, environmental, and public health concerns associated with conventional meat production. Cultured meat produced using cell culture technology promises to become a viable alternative to animal-raised meat for the future of the food industry. In this study, biomimetic bovine muscle tissue was artificially fabricated from myogenic cells extracted from bovine meat. Our primary culture method relies on three key factors; a sequential digesting process, enzymatic treatment with pronase, and coating with laminin fragment on culture dishes. This method allows the efficient collection of large numbers of primary cells from bovine cheek meat, purifies the myogenic cells from the cell mixture, and then continuously grows the myogenic cells in vitro. In addition, using our "quality control" methods, we were able to determine the "cell quality", including the proliferative and differentiation capability in each step of the primary culture. Furthermore, to mimic native bovine meat, the quality-controlled bovine myogenic cells were cultured on a micropatterned thermoresponsive substrate stimulating a native-like aligned structure of cells, which were then transferred onto a fibrin-based gel. This gel-based culture environment promoted structural and functional maturation of the myogenic cells, resulting in the production of bovine muscle tissues with sarcomere structures, native-like membrane structures, and contractile ability. We believe that these biomimetic features of "tissue-engineered meat" are important for the production of future cultured meat, which will need native-like nutrients, texture and taste. Therefore, our meat production approach will provide a new platform to produce more native biomimetic tissue-engineered meat in the near future.

Cultivated meat, also known as cell-based or clean meat, utilizes mesenchymal stem cells to cultivate mature cell types like adipocytes, which are pivotal for imparting the desired taste and texture. The delivery of differentiated cells, crucial in cultivated meat production, is facilitated through extensive exploration of 3D culturing techniques mimicking physiological environments. In this study, we investigated the adipogenic differentiation potential of bovine umbilical cord stem cells (BUSCs), sourced from discarded birth tissue, and assessed the feasibility of delivering differentiated cells for cultivated meat using gelatin methacrylate (GelMA) as a carrier for adipose tissue. Various adipogenic inducers, previously reported to be effective for human mesenchymal stem cells (hMSCs), were evaluated individually or in combination for their efficacy in promoting the adipogenesis of BUSCs. Surprisingly, while the traditional adipogenic inducers, including insulin, dexamethasone, isobutylmethylxantine (IBMX), indomethacin, and rosiglitazone, showed no significant effect on the adipogenic differentiation of BUSCs, efficient differentiation was achieved in the presence of a fatty acid cocktail. Furthermore, we explored methods for the delivery of BUSCs. Differentiated cells were delivered either encapsulated in GelMA hydrogel or populated on the surface of GelMA microparticles (MPs) as the adipose component of cultivated meat. Our findings reveal that after adipogenic induction, the lipid production per cell was comparable when cultured either within hydrogel or on MPs. However, GelMA-MPs supported better cell growth compared to hydrogel encapsulation. Consequently, the overall lipid production is higher when BUSCs are delivered via GelMA-MPs rather than encapsulation. This study not only systematically evaluated the impact of common adipogenic inducers on BUSCs, but also identified GelMA-MPs as a promising carrier for delivering bovine adipocytes for cultivated meat production.

Think about what humans will eat in the future. Could you imagine eating meat produced without the extensive farming of animals? This scenario is not as unlikely as you might think. To protect our planet, we must reduce the irreversible damage we are doing to the environment. Eating less meat is a major part of this, because the way we produce most meat today has a tremendous impact on the environment. Plant-based meat alternatives are already available in stores, but it is hard to imitate the taste and texture of meat if no animals are involved. Therefore, scientists have been working on an alternative, called cultivated meat, that is made from real animal tissue but does not require the death of the animal. Cultivated meat shows great promise to become an environmentally and animal-friendly alternative to conventional meat. However, there are still limitations to cultivated meat products that prevent them from being sold in stores.

The in vitro production of animal-derived foods via cellular agriculture is emerging as a key solution to global food security challenges. Here, the potential for fiber-based scaffolds, including silk and cotton, in the cultivation of muscle cells for tissue formation was pursued. Mechanical properties and cytocompatibility with the mouse myoblast cell line C2C12 and immortalized bovine muscle satellite cells (iBSCs) were assessed, as well as pre-digestion options for the materials due to their resilience within the human digestive track. The fibers supported cell adhesion, proliferation, and guided muscle cell orientation, facilitating myotube formation per differentiation. A progressive increase in biomass was also documented. Interestingly, iBSC proliferation was enhanced with coatings of recombinant proteins while C2C12 cells showed minimal response. Thus, both cotton and silk yarns were suitable as fiber-based scaffolds towards cell supportive goals, suggesting an alternative path toward structured protein-rich foods via this initial stage of textile engineering for food. Biomass prediction models were generated, enabling forecasts of cell growth and maturation across various scaffold conditions and cell types. This capability enhances the precision of the cultivation process towards an engineering approach, building on the inherent benefits of hierarchical muscle tissue structure, but here via textile engineering with these initial muscle-coated edible fibers. Further, the approach offers to reduce costs by optimizing cultivation time and media needs. These approaches are part of a foundation for future scalable and sustainable cultivated meat production. STATEMENT OF SIGNIFICANCE: This research investigates the use of one-dimensional fiber-based scaffolds for cultivated meat production, contributing to advancements in cellular agriculture. It introduces a method to measure changes in biomass and scaffold degradation throughout the cultivation process. Additionally, our development of biomass prediction models improves the precision and predictability of cultivated meat production. This research not only aids in scaling up cultivated meats but also enhances the use of textile engineering techniques in tissue engineering, paving the way for producing complex, three-dimensional meat structures more sustainably.

Cultivated meat (CM) refers to edible lab-grown meat that incorporates cultivated animal cells. It has the potential to address some issues associated with real meat (RM) production, including the ethical and environmental impact of animal farming, and health concerns. Recently, various biomanufacturing methods have been developed to attempt to recreate realistic meat in the laboratory. We therefore overview recent achievements and challenges in the production of CM. We also discuss the issues that need to be addressed and suggest additional recommendations and potential criteria to help to bridge the gap between CM and RM from an engineering standpoint.

This study compared the cellular and genetic characteristics of bovine skeletal muscle satellite cells (SMSCs) from Hanwoo (a Korean native cattle breed), including calves and mature cattle. SMSCs were isolated using magnetic-activated cell sorting (MACS) from tissue samples of six Hanwoo (three calves and three mature cattle) using the CD29 antibody. Calves’ SMSCs exhibited significantly faster growth rates than did those from cattle (P < 0.01), with a doubling time of 2.43 days. Genetic analysis revealed higher MyoD and Pax7 expression in SMSCs from calves during proliferation than in those from mature cattle (P < 0.001). However, FASN and PLAG1 expression levels were higher in mature cattle than in calves during both proliferation and differentiation (P < 0.001). These findings highlight the need for strategies to improve bovine muscle cell growth to produce competitive cultivated meat at a competitive price.

Cellular agriculture is an emerging research field of agribiotechnology that aims to produce agricultural products using stem cells, without sacrificing animals or cultivating crops. Cultivated meat, as a representative cellular product of cellular agriculture, is being actively researched due to global food insecurity, environmental, and ethical concerns. This review focuses on the application of stem cells, which are the seeds of cellular agriculture, for the production of cultivated meat, with emphasis on deriving and culturing muscle and adipose stem cells for imitating fresh meat. Establishing standards and safety regulations for culturing stem cells is crucial for the market entry of cultured muscle tissue-based biomaterials. Understanding stem cells is a prerequisite for creating reliable cultivated meat and other cellular agricultural biomaterials. The techniques and regulations from the cultivated meat industry could pave the way for new cellular agriculture industries in the future.

Scaffolds suitable for use in food products are essential in cultured meat production. Simultaneously, efforts are being undertaken to strengthen the scaffolding to improve cell proliferation, differentiation, and tissue formation. Muscle cells proliferate and differentiate according to the directional patterns of the scaffold, similar to natural tissue and native muscle tissue. Therefore, establishing an aligned pattern in the scaffolding architecture is vital for cultured meat applications. Recent studies on the fabrication of scaffolds with aligned porosity structures and their utility in manufacturing cultured meat are highlighted in this review. In addition, the directional growth of muscle cells in terms of proliferation and differentiation has also been explored, along with the aligned scaffolding architectures. The aligned porosity architecture of the scaffolds supports the texture and quality of meat-like structures. Although it is difficult to build adequate scaffolds for culturing meat manufactured from diverse biopolymers, it is necessary to develop novel methods to create aligned scaffolding structures. Furthermore, to avoid animal slaughter in the future, it will be imperative to adopt non-animal-based biomaterials, growth factors, and serum-free media conditions for quality meat production.

Global demand for animal protein is on the rise, but many practices common in conventional production are no longer scalable due to environmental impact, public health concerns, and fragility of food systems. For these reasons and more, a pressing need has arisen for sustainable, nutritious, and animal welfare-conscious sources of protein, spurring research dedicated to the production of cultivated meat. Meat mainly consists of muscle, fat, and connective tissue, all of which can be sourced and differentiated from pluripotent stem cells to resemble their nutritional values in muscle tissue. In this paper, we outline the approach that we took to derive bovine embryonic stem cell lines (bESCs) and to characterise them using FACS (fluorescence-activated cell sorting), real-time PCR and immunofluorescence staining. We show their cell growth profile and genetic stability and demonstrate their induced differentiation to mesoderm committed cells. In addition, we discuss our strategy for preparation of master and working cell banks, by which we can expand and grow cells in suspension in quantities suitable for mass production. Consequently, we demonstrate the potential benefits of harnessing bESCs in the production of cultivated meat.

The commercial growth factors (GFs) and serum proteins (SPs) contribute to the high cost associated with the serum-free media for cultivated meat production. Producing recombinant GFs and SPs in scale from microbial cell factories can reduce the cost of culture media. Escherichia coli is a frequently employed host in the expression recombinant GFs and SPs. This review explores critical strategies for cost reduction in GFs and SPs production, focusing on yield enhancement, product improvement, purification innovation, and process innovation. Firstly, the review discusses the use of fusion tags to increase the solubility and yield of GFs & SPs, highlighting various studies that have successfully employed these tags for yield enhancement. We then explore how tagging strategies can streamline and economize the purification process, further reducing production costs. Additionally, we address the challenge of low half-life in GFs and SPs and propose potential strategies that can enhance their stability. Furthermore, improvements in the E. coli chassis and cell engineering strategies are also described, with an emphasis on the key areas that can improve yield and identify areas for cost minimization. Finally, we discuss key bioprocessing areas which can facilitate easier scale-up, enhance yield, titer, and productivity, and ultimately lower long-term production costs. It is crucial to recognize that not all suggested approaches can be applied simultaneously, as their relevance varies with different GFs and SPs. However, integrating of multiple strategies is anticipated to yield a cumulative effect, significantly reducing production costs. This collective effort is expected to substantially decrease the price of cultivated meat, contributing to the broader goal of developing sustainable and affordable meat.

No abstract available

Cellular agriculture provides a potentially sustainable way of producing cultivated meat as an alternative protein source. In addition to muscle and connective tissue, fat is an important component of animal meat that contributes to taste, texture, tenderness, and nutritional profiles. However, while the biology of fat cells (adipocytes) is well studied, there is a lack of investigation on how adipocytes from agricultural species are isolated, produced, and incorporated as food constituents. Recently we compiled all protocols related to generation and analysis of adipose progenitors from bovine, porcine, chicken, other livestock and seafood species. In this review we summarize recent developments and present key scientific questions and challenges that need to be addressed in order to advance the biomanufacture of 'alternative fat'.

No abstract available

Global food waste has become an important environmental challenge. In this study, we established a hierarchical valorisation system through 'acidogenic fermentation-residue saccharification-photosynthetic bacteria (PSB) protein production'. Acidogenic self-fermentation for 72 h yielded liquid chemical oxygen demand (COD) and lactic acid levels of 56443.33 and 11634.64 mg/L, respectively. Solid-phase residues from the fermentation broth underwent enzymatic saccharification at 60 °C, yielding 68.7 % carbohydrate conversion and 16278 mg/L reducing sugar. The liquid phase was treated with PSB, which resulted in 95 %, 89.7 % and 66.6 % removal of lactic acid, COD and ammonium-nitrogen (NH4+-N), respectively, and PSB biomass and protein of 1356.5 and 415.1 mg/g. The relative abundance of Rhodopseudomonas was 59.04 %, with a metabolic shift from carbohydrate utilization to amino acid synthesis. The enhanced expression of the genes Rubisco and GAPDH strengthened energy metabolism and promoted PSB growth. Safety assessments identified potential allergens in the PSB protein; however, in vitro digestibility reached 77.28 %. Heavy metal content was 85 %-97 % below the food safety limits, thus confirming the safety of the resulting protein.

No abstract available

During the production of ethyl alcohol from food raw materials, a significant amount of by-products of microbiological synthesis is formed. Therefore, the issue of waste disposal and rational use of post-alcohol grain barda is a relevant area of research. The work was carried out at the Gorsky State Agrarian University, in the laboratories of the Faculty of Biotechnology. The article presents the results of using yeast for the biosynthesis of protein from post-alcohol grain barda. The protein content in the yeast biomass was determined to be 21.8 g / l. This indicator exceeded the concentration in the initial substrate (corn barda) by 9.6 g / l, which is associated with the partial introduction of nutrient salts and the accumulation of yeast biomass up to 967 million CFU / ml as a result of fermentation for 3-4 hours.

Overabundant agro-industrial side streams such as lactose-rich effluents from dairy activities offer multiple valorisation opportunities. In the present study, a food-grade mixed culture of bacteria and yeasts was tested under different operational conditions for the treatment and the valorisation of cheese whey permeate (CWP), the residue of whey protein recovery, into microbial protein (MP). Under continuous aerobic fermentation settings, the carbon-to-nitrogen (C/N) ratio showed little to no influence on the system performances and MP quality as compared to dilution rates (D), leading to a final protein content as high as 76%. Under high D values, instead, while biomass productivity increased, N-efficiency and protein content decreased. Unlike the bacterial community, the yeast one proved to be highly stable and less influenced by the increase of D. A preliminary estimate indicated that 2-11% of the future MP-based food production could be satisfied by only valorising lactose-rich dairy residues such as CWP.

Pretreatment methods play a pivotal role in the efficient breakdown of lignocellulosic biomass to produce highly digestible solids. Incorporating multistage or combined pretreatments provides increased efficiency and further carbohydrate depolymerization. Unlike other biomass feedstocks, ryegrass is a promising nutritional plant protein source with a protein content of 6–16%. In this study, protein extraction was combined with a mechanochemical pretreatment and biochemical fermentation stage to maximize the value of products from the system. To this end, the effectiveness of cold press, ball mill, and mild alkaline combined treatments on protein extraction was investigated, and the efficiency of enzymatic saccharification of the resulting solid material and its use as a medium for the oleaginous yeast Metschnikowia pulcherrima was explored. The cold press allowed the extraction of approximately 39% of the crude protein. Ball milling assisted with 5% Na2CO3 provided a drastic increase in the surface area and decrease in the particle size, albeit it did not significantly alter the structure in favor of enzymatic hydrolysis. A subsequent 0.5% NaOH pretreatment achieved enhanced fermentable sugar production with 45.6 g/L total sugars realized, notably 11-fold and 3.8-fold higher compared to untreated and mechanochemical-treated samples, respectively. M. pulcherrima efficiently metabolized all monosaccharides presented in supplemented ryegrass hydrolysates, yielding 0.20 (Y m/m) biomass with a 38.7% lipid content, similarly in a synthetic medium. Apart from being a lignocellulosic feedstock-based fermentable sugar production system serving the microbial lipid bioprocess, cold-press integrated mechanochemical biorefinery was shown to be a promising approach in the extraction of plant protein, while preventing robust separation operations before the severe chemical biorefinery stages of ryegrass.

The conversion of nonedible biomass to protein for use in feed is an attractive strategy toward improved sustainability in aquaculture. We have studied the possibility to produce protein-rich yeast Candida utilis on a medium consisting of enzymatically hydrolyzed sulphite-pulped spruce wood, mainly providing glucose, and enzymatically hydrolyzed brown seaweed, supplemented with ammonium sulfate. The results show that this blend constitutes a complete fermentation medium that enables good growth rates and cell yields. Results from a salmon feeding trial showed that the yeast can replace parts of a traditional fishmeal diet without harmful effects, although the apparent protein digestibility coefficient for the yeast was suboptimal. While further optimization of both the fermentation process and downstream processing is needed, the present proof-of-concept study shows a path to the production of microbial protein based on a simple, local and sustainable fermentation medium.

This study investigated the quality evolution of soybean products (soymilk, tofu, dried bean curd) through mixed-strain fermentation with Lacticaseibacillus rhamnosus CICC 6151 and Saccharomyces cerevisiae AS2.400 under optimized conditions (7% inoculum, pH of 5.2, 85 °C/50 min thermal treatment). Physicochemical, structural, and microbial dynamics were systematically analyzed. Key results demonstrated that probiotic tofu exhibited superior water-holding capacity (82% WHC vs. 65% in traditional variants) and enhanced protein retention (Δ + 2.4% during storage), linked to microbial-mediated structural stabilization. Mixed fermentation induced substrate competition (S. cerevisiae biomass: OD560 of 1.2 at 10 h vs. L. rhamnosus OD600 of 1.0 at 25 h; ANOVA p < 0.001), driving pH-dependent protein network formation (isoelectric precipitation at pH of 4.8 ± 0.1) and volatile profile divergence (PCA explained 82.2–89.1% of variance). Probiotic variants maintained chromatic stability (ΔE < 15 vs. traditional ΔE > 23) and textural integrity (23% lower deformation under compression), correlated with secondary structure preservation (β-sheet increased by 10% in FTIR analysis). These findings establish synergistic microbial–metabolic regulation as a strategy for developing functional bean products with enhanced nutritional and sensory properties.

Fermented foods are staples in diets worldwide and are known for their health benefits. Microorganisms are the key to fermented food production as they convert raw substrates into digestible, nutritious, and health-promoting products. While microbes are essential for fermented food production, their contribution to the dietary protein profile of the final food product in terms of microbial biomass is largely unknown. We analyzed proteins from 17 fermented foods using metaproteomics to identify and quantify microbial and food-derived proteins. We found that microbial proteins contribute up to 11% of the total protein content in fermented foods and comprise as much as 60% of the total number of identified proteins. These microbial proteins included many for central functions in microbial cells, such as glycolysis enzymes, translation machinery, and chaperones, as well as proteins for specialized functions that are important for the ecological niches in food fermentation, such as carbohydrate degrading enzymes and proteases. Some of these microbial proteins, such as proteases, could impact gut physiology. These findings highlight the substantial contribution of microbial proteins to the nutritional and functional profile of fermented foods, which may have important implications for interactions with the gut microbiota and health outcomes.

The demand for cheap, healthy, and sustainable alternative protein sources has turned research interest into microbial proteins. Mycoproteins prevail due to their quite balanced amino acid profile, low carbon footprint and high sustainability potential. The goal of this research was to investigate the capability of Pleurotus ostreatus to metabolize the main sugars of agro-industrial side streams, such as aspen wood chips hydrolysate, to produce high-value protein with low cost. Our results indicate that P. ostreatus LGAM 1123 could be cultivated both in a C-6 (glucose)- and C-5(xylose)-sugar-containing medium for mycoprotein production. A mixture of glucose and xylose was found to be ideal for biomass production with high protein content and rich amino acid profile. P. ostreatus LGAM 1123 cultivation in a 4 L stirred-tank bioreactor using aspen hydrolysate was achieved with 25.0 ± 3.4 g L−1 biomass production, 1.8 ± 0.4 d−1 specific growth rate and a protein yield of 54.5 ± 0.5% (g/100 g sugars). PCA analysis of the amino acids revealed a strong correlation between the amino acid composition of the protein produced and the ratios of glucose and xylose in the culture medium. The production of high-nutrient mycoprotein by submerged fermentation of the edible fungus P. ostreatus using agro-industrial hydrolysates is a promising bioprocess in the food and feed industry.

No abstract available

Agricultural and industrial residues are renewable biomass sources present in large quantities causing pollution. Therefore, transforming these residues to eco-friendly products such as enzymes and bioactive materials reduces their quantity and impact on the environment, in addition to reducing the production costs. Sesame cake is a by-product of the production of Sesame seed oil and is high in protein. The yield of Sesame cake protein hydrolysis (SH) improved by 4.2-fold through the optimization of conditions using Bacillus thuringiensis strain-MA8 protease via the Box-Behnken design (BBd). The average diameter of the particle size of SH was 677.10 nm. The application of SH (1–3%) in the production of low-fat yogurt (LSH) exhibited a fermentation time similar to that enriched with skim milk powder (LSMP). The total solids and protein levels in LSH-yogurt exceeded those in full fat yogurt (FFY). In addition, the acidity and overall acceptability ratings of LSH-yogurt were similar to FFY throughout the 15-day storage at 5 °C, without displaying any defects. Furthermore, the total essential amino acids (TEAA), total amino acids (TAA), and TEAA/TAA ratio of LSH (2%)-yogurt were approximately similar to FFY. Incorporating SH (2%) improved the chemical score of certain amino acids in LSH-yogurt. The hardness of LSH-yogurt exceeded that of FFY. Additionally, the springiness, gumminess, and cohesiveness of LSH-yogurt were similar to those of LSMP. Protein hydrolysate from Sesame cake is a new fat substitute for low-fat yogurt production without displaying any defects as well as reducing the risks associated with high-fat consumption and global obesity.

This study evaluated Lolium perenne press juice as a sustainable substrate for Single-Cell Protein (SCP) production using Kluyveromyces marxianus. Key fermentation parameters were systematically optimized, including microbial reduction, dilution ratios, temperature, and nutrient supplementation. Pasteurization at 75 °C preserved essential nutrients better than autoclaving, resulting in a 27.8% increase in biomass yield. A 1:2 dilution of press juice enhanced fermentation efficiency, achieving 20.2% higher biomass despite a lower initial sugar content. Cultivation at 30 °C enabled sustained substrate utilization and outperformed 40 °C fermentation, increasing final biomass by 43.4%. Nutrient supplementation with yeast extract, peptone, and glucose led to the highest biomass yield, with a 71% increase compared to unsupplemented juice. Press juice from the tetraploid variety, Explosion, consistently outperformed the diploid Honroso, especially when harvested early, reaching up to 16.62 g·L−1 biomass. Early harvests promoted faster growth, while late harvests exhibited higher biomass yield coefficients due to improved sugar-to-biomass conversion. Compared to a conventional YM medium, fermentation with L. perenne press juice achieved up to a threefold increase in biomass yield. These findings highlight the potential of grass-based substrates for efficient SCP production and demonstrate how agricultural parameters like variety and harvest timing influence both quantity and quality. The approach supports circular bioeconomy strategies by valorising underutilized biomass through microbial fermentation.

Study’s Excerpt: Aspergillus niger was used to produce single-cell protein from yam peel waste. Yam peels showed high carbohydrate content (81.73%) and supported fungal growth. Biomass yield was higher on yam peel (0.4 OD) than on commercial YEPD medium (0.23 OD). Yam peel substrate proved a viable low-cost medium for SCP production. The study promotes SCP production as a use for agro-waste and pollution control. Full Abstract: Agro-industrial waste is a source of nutrients and compounds that can be used to support microbial growth in fermentation processes for the production of bio-products such as enzymes, antibiotics, or single-cell proteins. The microbial biomass is a source of proteins with several advantages over traditional protein sources. This study aimed to produce single-cell protein (SCP) from Aspergillus niger using yam peels as substrate. Aspergillus niger was isolated from garden soil using Sabouraud dextrose agar (SDA) and characterized accordingly. Yam peels a restaurant in processed and analyzed for proximate composition according to standard protocols. The isolate was subjected to submerged fermentation using commercially prepared yeast extract peptone dextrose (YEPD), and the yam peel substrate for a period of 7days on a rotary shaker. Results show that Aspergillus niger isolates had dark to brown colonies with black conidial heads and a pale yellow colour on the reverse of the SDA plate. Microscopically, the conidiophore extended from its hyphae, carrying black globular conidia.  Proximate composition of the yam peels substrate was found to contain carbohydrates (81.73%), lipids (4.17%), proteins (3.5%), moisture (5.19%), ash (5.4%), and fiber (1.85%). Support for higher fungal biomass was observed on yam peel substrate, which attained 0.4 OD (optical density), while the maximum growth on the commercially prepared media (YEPD) was 0.23 OD. Thus, the Yam peel substrate supported significantly higher A. niger biomass yield (0.4 OD) compared to commercial YEPD medium (0.23 OD). It is recommended that agro-industrial wastes such as yam peels and related wastes be used to enhance production of SCP, thereby reducing pollution caused by improper disposal of agro-industrial wastes.

The growing global population and climate crisis demand expanding non-animal protein options. Single-cell protein biomass, referred to as "Solein", is produced by the hydrogen-oxidising bacterium Xanthobacter sp. SoF1 and is a promising, sustainable source of protein and dietary fibre, especially when created using renewable energy. This study investigates Solein protein powder (SPP) for its composition and techno-functional properties, comparing it to pea protein isolate (PPI). SPP had a lower fat content and higher dietary fibre, while matching the protein content of PPI. SPP met all indispensable amino acid requirements for adults over the age of three, as outlined by the FAO in 2013. A milk alternative resembling semi-skimmed cow's milk was produced from SPP and PPI. These emulsions were fermented with a commercial starter culture containing Streptococcus thermophilus. The fermentation process was monitored by tracking pH, total titratable acidity, and microbial growth. The resulting yoghurt alternative (YA) underwent textural and rheological analysis. Solein protein powder yoghurt alternative (SPP-YA) exhibited faster acidification, greater microbial growth, improved water retention, and a texture similar to dairy yoghurt. Static in vitro digestion revealed moderate protein digestibility of the non-fermented SPP emulsion (63.8-67.5%), based on total amino acids, free amino groups, and total nitrogen, with an in vitro Digestible Indispensable Amino Acid Score (DIAAS) of (51.0 ± 6.1%). Fermentation slightly reduced digestibility (57.8-59.6%) and DIAAS (48.3 ± 1.4%), with isoleucine as the limiting amino acid. This work provides the first insight into the structural and nutritional performance of hydrogen-oxidising bacterial protein in non-dairy YA.

No abstract available

Caragana korshinskii Kom. represents a substantial biomass resource that can be converted into feed protein via microbial fermentation. This study aimed to improve the nutritional value of C. korshinskii through strain screening and substrate optimization. Amino acid content and in vitro digestibility were systematically investigated. Astral-DIA proteomics was employed to compare protein enrichment mechanisms underlying screened microbial involvement in substance conversion. The Aspergillus oryzae and Saccharomyces cerevisiae co-culture increased the true protein content of the optimized substrate by 50.6% to 67.9%, while the highest nitrogen conversion ratio (69.5%) was achieved with low-level supplementation of (NH4)2SO4. The relative abundances of hydroxyproline and lysine content increased by more than twice in the mixed fermentation. Proteomics analysis identified 291 differentially expressed proteins in the mixed culture versus A. oryzae alone, enriched in ribosome biogenesis; valine, leucine and isoleucine biosynthesis; galactose metabolism; amino acids biosynthesis and sulfur relay system. This study provided guidance for the high-value utilization of C. korshinskii and elucidated the differential protein enrichment pathways between A. oryzae, S. cerevisiae and their cocktail in utilizing C. korshinskii.

Rice straw and cassava waste are abundant agricultural byproducts, but they have limitations in nutritional value, particularly low protein content. This study aims to evaluate the effect of a combination of straw and cassava substrates, as well as the addition of Saccharomyces cerevisiae mold, on increasing the content of single cell protein (SCP), crude protein, pure protein, microbial biomass, and pH stability during fermentation. The study was conducted experimentally with a 2×2 factorial completely randomized design (CRD) using four treatments: JKN (straw + cassava peel without yeast), JKR (straw + cassava peel + yeast), JSN (straw + cassava without yeast), and JSR (straw + cassava + yeast). Fermentation was carried out for 7 days under anaerobic conditions. The results showed that the JSR treatment produced the highest SCP (47.59 mg/mL) and the largest microbial biomass (4.37 mg/mL), while JSN produced the highest crude protein (13,40%) and pure protein (9,61%). The addition of yeast significantly increased PST and biomass, but in some treatments, it decreased the pure protein content. The combination of straw and cassava has been shown to increase the nutritional value of feed ingredients through the fermentation process, with effectiveness depending on the type of substrate and the presence of microbial inoculum.

The growing need for high-quality protein with minimal environmental impact necessitates the expansion of alternative proteins on the market. One area with great opportunity for expansion lies in the phylogenetic diversity of the fungal kingdom. Diversifying the use of fungal species, by assessing species from the phylum of mushroom-producing fungi (Basidiomycota) in solid-state fermentation, could open new avenues to foods with improved nutritional and sensorial properties. To assess these properties, we first determined the potential of basidiomycetes to ferment and colonize cereals and legumes. A phylogenetically diverse selection of eight species of basidiomycetes was analyzed on their radial growth speed and biomass yield. The best performing species were successfully fermented on brown rice (high starch), brewer's spent grain (high fiber, high protein), and lupin (high protein, high fiber and high fat), and compared to Rhizopus microsporus var. oligosporus. Large variation in performance was observed between the different basidiomycetes on the three substrates in terms of biomass formation and metabolic behavior. The presence of an easily accessible carbon source, such as starch was needed to prevent deamination and thereby loss of valuable protein. With the correct formulation, basidiomycetes could fully ferment and colonize the substrate, thereby increasing the overall protein content and degrading the anti-nutritional factor phytic acid up to 80 %. These results provide a methodology for screening of fungal species and substrates and demonstrate that basidiomycetous mycelia represent a promising source of phylogenetic diversity for novel food fermentations.

No abstract available

Innovative protein sources are urgently needed to feed a growing global population and to support the increasing shift toward vegetarian and vegan lifestyles. Mycelia of edible fungi offer a sustainable and efficient alternative food source. In this study, 106 fungal strains were explored for their ability to ferment two different liquid carrot side streams. Among the candidates, Pleurotus djamor demonstrated exceptional potential, with high yields of biomass of ∼15 g L–1 and high protein contents of 31.0 ± 5.9 (optimized orange carrot medium) or 21.6 ± 1.9 g 100 g–1 (optimized black carrot medium), respectively. When used in burger patties and vegan sausage analogs, the mycelia outperformed vegetable proteins in sensory tests, highlighting their viability as a nutritious, versatile, and consumer-accepted protein alternative.

ABSTRACT The aim of this study was to hydrolyze cultivated fungal mycelium and to evaluate the effect on its taste. Potato pulp, a by‐product of the potato starch industry, was therefore successfully utilized as a substrate for submerged cultivation of Flammulina velutipes, yielding a product with an estimated fungal content of 83% ± 3%. The fermentation increased the protein content from 5.3 ± 0.4 g/100 g DM to 13.9 ± 0.1 g/100 g DM with a biological protein value of 86. The fermentate was enzymatically hydrolyzed by Corolase APC‐peptidase. After optimization of the hydrolysis conditions, a degree of hydrolysis (DH) of 75.1% ± 1.0% was achieved. The protein hydrolysis increased the contents of free glutamate more than 20‐fold from 8.7 ± 0.1 mg/L to 188.7 ± 1.2 mg/L. Elevated glutamate levels led to an umami taste perception in aqueous solution and taste‐enhancing properties in vegetable broth. Noteworthy, the fermentate itself exhibited an intrinsic peptidase activity. Without addition of auxiliary peptidases, mycelial enzymes caused a DH of 33.9% ± 0.7% and a free glutamate content of 99.1 ± 0.7 mg/L. For these samples, an increase in umami taste was only observed in vegetable broth, but not in water, indicating taste‐enhancing properties but low umami taste. In addition to the nutritional and health benefits of fungi, their hydrolysates are of great interest for use as a protein booster with flavor‐enhancing properties.

Food production is one of the most environmentally damaging human activities. In the face of climate change, it is essential to rethink our dietary habits and explore potential alternative foods catering both towards human and planetary needs. Fungal mycelium might be an attractive alternative protein source due to its rapid growth on sustainable substrates as well as promising nutritional and organoleptic properties. The natural biodiversity of filamentous fungi is vast and represents an untapped reservoir for food innovation. However, fungi are known to produce bioactive compounds that may affect human health, both positively and negatively. To narrow the search for safe and culinarily attractive fungal species, mycelia of edible fruiting-body forming fungi provide a promising starting point. Here, we explore whether the culinary attractiveness and safety of the commonly eaten mushroom, Pleurotus ostreatus, can also be translated to its mycelium. Whole-genome sequencing and pan-genome analysis revealed a high degree of genetic variability within the genus Pleurotus, suggesting that gastronomic traits as well as food safety may differ between strains. A representative strain, P. ostreatus M2191, was further analyzed for the food safety, nutritional properties and culinary applicability of its mycelium. No regulated mycotoxins were detected in either the fruiting body nor the mycelium. Yet, P. ostreatus is known to produce four peptide toxins, Ostreatin, Ostreolysin and Pleurotoysin A/B. These were found to be lower in the mycelium compared to fruiting bodies, which are already considered safe for consumption. Instead, a number of secondary metabolites with potential health benefits were detected in the fungal mycelium. In silico analysis of the proteome suggested low allergenicity. In addition, the fruiting body and the mycelium showed similar nutritional value, which was dependent on the growth substrate. To highlight the culinary potential of mycelium, we created a dish served at the two-star restaurant the Alchemist in Copenhagen, Denmark. Sensory analysis of the mycelium dish by an untrained consumer panel indicated consumer liking and openness to fungal mycelia. Based on sustainability, safety, culinary potential, and consumer acceptance, our findings suggest that P. ostreatus mycelium has great potential for use as a novel food source.

Abstract Cocoa pod husks (CPHs), the major side‐stream from cocoa production, were valorized through fermentation with Pleurotus salmoneo‐stramineus (PSS). Considering ergosterol as a biomarker for the fungal content, the mycelium accounted for 54% of the total biomass after 8 days in submerged cultures. The crude protein content of fermented CPH (CPHF) increased from 7.3 g/100 g DM in CPH to 18.9 g/100 g DM. CPH fermentation resulted in a high biological value of 86 for the protein. The water and oil binding capacities of CPHF were 3.5 mL/g and 2.1 mL/g, respectively. The particle diameter dv,0,90 of CPHF was 373 μm as compared to 526 μm for CPH. The total dietary fiber was 73.4 g/100 g DM in CPHF and 63.6 g/100 g DM in CPH. The amount of soluble fiber was 2.3 g/100 g DM in CPHF and 10.1 g/100 g DM in CPH; the insoluble fraction accounted for 71.1 g/100 g DM and 53.6 g/100 g DM, respectively. Bread doughs with CPH or CPHF were characterized for texture, color, and farinographic properties. The dough hardness, consistency, and browning index increased with the concentration of CPH, whereas for CPHF, springiness and peak viscosities declined. We demonstrate the upcycling of CPH into nutritious and functional ingredients through PSS fermentation.

This study investigated the effects of ultrasonic treatment with different intensities (200, 400, and 600 W) and durations (15, 30, and 45 min) on the mycelial integrity, product quality, and in-vitro digestibility of mycoprotein. Results showed that the mycelium was damaged to varying degrees under different ultrasonic parameters. The mycelium showed obvious damage when the time of ultrasonic treatment lasted more than 30 min or the power exceeded 400 W. As a result, the particle size, hardness and chewiness of mycoprotein products were decreased significantly. Compared with the control group (CK), ultrasonic treatment increased the proportion of immobile water and changed the secondary structure of the fungal intracellular protein, which may be the reason for improving in-vitro digestibility of mycoprotein products. This result demonstrated that, by damaging the mycelial integrity and changing the protein secondary structure, ultrasound treatment could decrease the texture characteristics, but improve the water-holding capacity and in-vitro digestibility of products. This study would provide a potential application for the minced meat products by mycoprotein treated with ultrasound.

No abstract available

The growing aquaculture industry has an increasing demand for novel, sustainably produced protein sources for aquafeed. This study aimed to determine the apparent digestibility (AD%), pellet quality, and protein score of four novel fungal proteins in rainbow trout (Oncorhynchus mykiss), namely, PEKILO® (PEK) derived from Paecilomyces variotii, Aspergillus oryzae (AO), Rhizopus oligosporus (RO), and Rhizopus delemar (RD). All fungi were grown on various side-streams, such as beet vinasse, thin stillage, and whole stillage. The diets were produced by extrusion technology and consisted of control and test diets with a 30:70 test ingredient/control ratio. Feeding lasted for 39 days. Each tank had 20 fish, with three replicates per dietary treatment. One-way ANOVA was performed to compare the means of the groups with each other. The dry matter (DM) digestibility of PEK was significantly higher than that of AO, RD, and RO, all with similar digestibility. The crude protein AD% for PEK was 86.5%, which is significantly higher than that of the other fungal sources. AO, PEK, RD, and RO had similar crude fat AD% compared to each other, at 83.8%, 87.4%, 90.5%, and 88.5%, respectively. The pellet quality was found to deteriorate with addition of fungal proteins. PEK had high AD% for most of the macronutrients tested and better pellet quality.

No abstract available

No abstract available

Cheese whey is an industrial by-product that is generated in excess during the cheese production process in the dairy industry. Despite the potential utility of whey, it continues to pose environmental threats in the industry. This study comprehensively evaluates the utilization of two fermentation techniques (solid-state fermentation and submerged fermentation) for producing fungal biomass from cheese whey powder, employing Aspergillus oryzae, Rhizopus oryzae, and Neurospora intermedia for sustainable food production. It has been observed that submerged fermentation is more effective in increasing the protein content of whey powder compared to solid-state fermentation. The highest biomass yield was achieved with A. oryzae (5.29 g/L, 0.176 g biomass/g substrate), followed by N. intermedia (3.63 g/L, 0.121 g biomass/g substrate), and R. oryzae (1.9 g/L, 0.063 g biomass/g substrate). In the bubble column reactor, the protein content of the substrate (78.65 g/kg) increased by 165.54 and 176.69% with A. oryzae (208.85 g/kg) and N. intermedia (217.62 g/kg), respectively. This study has demonstrated that whey powder can be converted into protein-rich biomass through fungal bioconversion. The obtained biomass has the potential to be developed as an alternative food and feed source, contributing to waste management and sustainable food production.

No abstract available

Dysphagia represents a significant public health concern, and there is an increasing demand for the development of food products tailored to the needs of dysphagia patients. Therefore, this study employed Pleurotus ostreatus mycelium (POM), xanthan gum (XG), and egg white protein (EWP) as raw materials to fabricate dietary products suitable for dysphagia patients using three‐dimensional (3D) printing technology. The study systematically investigated the impact of varying POM concentrations (130, 150, 170, 190, and 210 g kg−1) on the physicochemical properties, structural characteristics, and textural attributes of 3D‐printed food matrices.

The growing demand for a more sustainable and nutritious food supply has increased interest in replacing animal-derived foods with those from alternative sources, such as fermentation processes. However, creating foods entirely from ingredients generated using a single fermentation process is often challenging. Consequently, there is interest in combining different sources of fermentation-derived ingredients to create foods with improved physicochemical, sensory, and nutritional properties. In this study, we examined the potential of combining two functional ingredients obtained using microbial fermentation. Mycoprotein (MCP) is a protein-rich material derived from mycelium fermentation that can form fibrous meat-like structures and has good nutritional properties, but it has poor gelling properties, which limits its ability to create meat substitutes and analogs. High acyl gellan gum (HA-GG) is a polysaccharide derived from bacterial fermentation that has excellent gelling properties. We therefore combined MCP and HA-GG to create hybrid hydrogels suitable for formulating meat substitutes and analogs. Differential scanning calorimetry, dynamic shear rheology, texture profile analysis, and scanning electron microscopy were used to assess the thermal, rheological, textural, and structural properties of MCP + HA-GG hydrogels with different compositions. The pure MCP (10 w/w%) samples did not exhibit any strong thermal transitions when heated or cooled from 10 to 90 °C. In contrast, pure HA-GG (2 w/w%) melted when heated above 85 °C and gelled when cooled below 80 °C. The MCP + HA-GG hybrids maintained a high shear modulus during both heating and cooling, which may be useful for food applications. The hybrids had an appreciably higher gel strength (Young's modulus, hardness, and shear modulus) than the individual MCP or HA-GG samples, which was attributed to a synergistic interaction between these two components. The gel strength, breaking stress, and breaking strain increased with increasing gellan gum concentration (0.5 to 2.0 w/w%), which meant that the mechanical properties of the hybrid materials could be tailored for specific applications. This study highlights the complex interactions among ingredients from alternative sources and their significant impact on the properties of food matrices. This information may be useful for formulating meat substitutes and analogs with enhanced physicochemical and functional properties.

There is increasing interest in the development of meat analogs due to growing concerns about the environmental, ethical, and health impacts of livestock production and consumption. Among non-meat protein sources, mycoproteins derived from fungal fermentation are emerging as promising meat alternatives because of their natural fibrous structure, high nutritional content, and low environmental impact. However, their poor gelling properties limit their application in creating meat analogs. This study investigated the potential of creating meat analogs by combining mycoprotein (MCP), a mycelium-based protein, with potato protein (PP), a plant-based protein, to create hybrid products with meat-like structures and textures. The PP-MCP composites were evaluated for their physicochemical, rheological, textural, and microstructural properties using electrophoresis, differential scanning calorimetry, dynamic shear rheology, texture profile analysis, confocal fluorescence microscopy, and scanning electron microscopy analyses. The PP-MCP hybrid gels were stronger and had more fibrous structures than simple PP gels, which was mainly attributed to the presence of hyphae fibers in mycelia. Dynamic shear rheology showed that the PP-MCP hybrids formed irreversible heat-set gels with a setting temperature of around 70 °C during heating, which was attributed to the unfolding and aggregation of the potato proteins. Confocal and electron microscopy analyses showed that the hybrid gels contained a network of mycelia fibers embedded within a potato protein matrix. The hardness of the PP-MCP composites could be increased by raising the potato protein content. These findings suggest that PP-MCP composites may be useful for the development of meat analogs with more meat-like structures and textures.

Dietary intervention is the basis for the treatment of diabetes mellitus. This study employed Ganoderma lucidum (GL) mycelium to ferment a compound medium of oat and purple potato (OPP), optimized fermentation conditions to increase the triterpene content in the resulting product (F-OPPF), and systematically investigated the impact of fermentation on the nutritional quality, structural characteristics, and functional properties of OPP. The results indicated that the triterpene content in F-OPPF significantly increased from 8.53 mg/g to 17.23 mg/g under optimal conditions (temperature: 28 °C, inoculum size: 10%, material quantity: 36 g/250 mL, and fermentation time: day 13). Fermentation resulted in enhanced nutritional quality, with increased contents of protein, soluble protein, crude fiber, ash, mineral elements, essential amino acids, polysaccharides, flavonoids, and total phenols. Mycelium not only enveloped the OPP surface but also penetrated its interior, forming a porous honeycomb-like structure. The types of reactive groups and crystals (C + V-type) were not changed after fermentation, while the crystallinity increased. F-OPPF exhibited positive changes in thermogravimetric properties, antioxidant and hypoglycemic activities, and adsorption capacity of insoluble dietary fiber. Additionally, incorporating F-OPPF into the diet markedly reduced fasting blood glucose levels and promoted weight gain in T2DM rats induced by a high-fat diet and streptozotocin. The fermented groups exhibited improvements in glyco- and lipo-metabolism, oxidative stress, and the function and pathological morphology of the pancreas, liver, and kidneys compared to the unfermented group. Collectively, these findings suggested that GL mycelium fermentation enhanced the nutritional and functional values of OPP, and F-OPPF holds potential as a raw material for developing diabetic-friendly foods.

This study presents a green, integrated strategy for valorizing fungal mycelium from Phellinus linteus (PL) and Cordyceps sinensis (CS) into functional yogurt alternatives. The effects of high-pressure homogenization (HPH), thermal pasteurization, and microbial fermentation on structural constituents and rheological properties were systematically investigated. HPH reduced particle size (by 77% in PL, 67% in CS) and increased protein solubility (35.7% in PL, 42.7% in CS), promoting the transition from a liquid-like system to a viscoelastic network. Prolonged pasteurization (85 °C, 30 min) enhanced starch gelatinization compared to shorter treatment (5 min), improving viscosity, water-binding capacity, and microbial safety. Fermentation with commercial yogurt starters reached pH 4.5 within 7 h and probiotic levels of 8-9 log colony-forming units (CFU)/g. In situ production of dextran and β-glucan by Weissella confusa VIII40 and Pediococcus claussenii 55 T, respectively, significantly enhanced viscosity, viscoelastic moduli, and creep-recovery behavior. Notably, high dextran yields (2.6-3.1%) significantly reduced syneresis to below 10%. Overall, the integrated mechanical and microbial processing demonstrates a clean-label method for converting underutilized fungal biomass into high fiber yogurt alternatives with improved rheological and textural quality.

No abstract available

No abstract available

Precision fermentation is emerging as an innovative platform for manufacturing high-value food proteins, relocating food production from agricultural fields to controlled bioreactors. Importantly, these proteins must preserve the precise amino acid composition and structural properties that underpin the functionality, texture, and nutritional value of their animal-derived counterparts. However, bulk food proteins are high-volume, low-value products and therefore need to be produced at scale as cheaply as possible. Currently, intracellular protein expression requires costly cell-lysis and downstream purification steps, which comprise product purity and generally result in the product not being cost-competitive with conventional agriculture. Thus, the commercial viability of lab-grown food proteins including animal-free dairy, egg, and collagen hinges on the capacity of microbial hosts, primarily yeasts and filamentous fungi, to export correctly folded proteins into the culture medium at gram-per-liter titers, in a process known as protein secretion. Yeast and fungi are ideal host organisms due to their potential for high-yield secretion and ability to reproduce many eukaryotic post-translational modifications. Accordingly, the protein secretory pathway now sits at the crucial intersection of synthetic biology, protein engineering, and bioprocess optimization. This perspective will address modifications to the secretory pathway that can improve protein secretion efficiency. Deliberate, data-driven engineering of secretion efficiency will determine whether precision-fermented proteins advance from pilot production to routine industrial manufacture. This perspective will also address the issues of cost efficiency and scalability, exploring how protein secretion can overcome these challenges to make lab-grown food a sustainable, ethical, and viable alternative to conventional food sources.

Dehulled peas (DHP) are increasingly popular in food applications, but their integration in food products is limited due to high starch and fiber causing gelling and aggregation during cooking. Additionally, DHP contains saponins (secondary metabolite), contributing to bitter flavors, yet they also hold health benefits. We hypothesize that fungal fermentation could enhance DHP nutritional profile and integration into food products. This study evaluated the effects of six fungal organisms (Aspergillus niger [An], Aspergillus oryzae [Ao], Aureobasidium pullulans [Ap], Neurospora crassa [Nc], Rhizopus microspores var. oligosporus [Ro], Trichoderma reesei [Tr]) on DHP over 120 h of submerged fermentation, evaluating total phenolics, starch, saponins, crude proteins, and overall mass balance. Results from the study demonstrated notable changes post‐fermentation, including increased overall protein content and solubility, decreased starch content, reduced overall mass recovery, and elevated levels of total phenolics and saponins. Filamentous fungi exhibited a significant reduction in starch content, contributing to a substantial reduction in mass recovery (31%–60%) compared to the control. Unexpectedly, saponin concentrations increased (1.5 to 3 folds) during fermentation, possibly attributed to the breakdown of the substrate matrix and release of bound saponins. Total phenolic levels varied among microorganisms, with An and Nc demonstrating the highest increases (6 to 10 folds) as compared to the control. Overall, these findings point to fungal fermentation as a tool for adding value to yellow peas and other crops facing similar processing challenges. Further research is warranted to understand the health impacts and value of these enhancements.

The protein content of a plant-based ingredient is generally lower than its animal food counterpart, and research into novel alternative protein is required that can provide similar protein content, texture and appearance as meat. This work investigates a mycelium-based low moisture meat analogue (LMMA) approach, by incorporating 0 to 40% w/w mycelium (MY) into pea protein isolate (PPI) via extrusion using a twin-screw extruder at 140 °C die temperature, 40 rpm screw speed, and 10 rpm feeder speed (0.53–0.54 kg/h). Physicochemical, rehydration, and structural properties of LMMA were assessed. The MY incorporation led to a significant change in color attributes due to Maillard reaction during extrusion. Water solubility index and water absorption capacity increased significantly with MY addition, owing to its porous structure. Oil absorption capacity increased due to increased hydrophobic interactions post-extrusion. Protein solubility decreased initially (upto 20% w/w MY), and increased afterwards, while the water holding capacity (WHC) and volumetric expansion ratio (VER) of LMMA enhanced with MY addition upto 30% w/w. Conversely, WHC and VER decreased for 40% w/w which was verified with the microstructure and FTIR analysis. Overall, MY (30% w/w) in PPI produced a fibrous and porous LMMA, showing future potential with an increasingly plant-based product market and decreasing carbon footprint of food production activities.

No abstract available

This research was conducted by utilizing anaerobic digested manure wastewater (ADMW) as a growth medium for microalgae Euglena sp. IDN 22. This research aimed to determine the effectiveness of using ADMW as a rich nutrient supplement to cultivate microalgae Euglena sp. IDN 22. Multiple dilutions of 0%, 25%, 50%, and 75% were applied to ADMW, and the growth of Euglena sp. IDN 22 was analyzed using the growth rate of microalgae cell counts. The number of cells was observed with a haemocytometer. The cultivation of Euglena sp. IDN 22 in the ADMW medium was conducted in lab-scale (1 L) photobioreactors for 15 days of cultivation. The analysis of 25% ADMW showed C/N and P/N of 7.40 ± 0.17 and 1.27± 0.03, while 50% ADMW analysis yielded 14.99 ± 0.51 and 1.72 ± 0.07. The best growth rate trend of Euglena sp. IDN 22 in ADMW was shown by the treatment with 25% ADMW dilution with a maximum growth of 62.92 × 10 4 cells/ml, compared to a maximum growth of 79.92 × 10 4 cells/ml with 0% ADMW as a control. Slow growth activity was shown by 50% and 75% ADMW, with a maximum growth of only 10.67 × 10 4 cells/ml and 4.45 × 10 4 cells/ml, respectively. The slow growth activity in 50% ADMW, with a maximum growth of only 10.67 × 10 4 cells/ml, demonstrated a significant difference (p > 0.01). The results of this study prove that the integration of 25% ADMW and Euglena sp. IDN 22 cultivation can be an effective valorization concept and an opportunity to transform biogas production waste towards a circular economy in agroindustry.

This study aimed at developing a sustainable waste management from poultry farm by integrating microalgae cultivation with the anaerobic digestion effluent of chicken wastes (ADECW). The analysis was focused on system performance, resource recovery and environmental impact of microalgal biomass-derived added value products. Laboratory-scale of three different systems, i.e. suspended microalgae, biofilm microalgae and the control as no microalgae seed added, was conducted under outdoor climatic conditions in Thailand. The results clearly showed that microalgae system was successfully developed with high treatment performance and potential renewable energy production for the ADECW. Compared to the control, it was demonstrated that most removals of nutrient and organic pollutants were achieved through microalgal assimilation. Biofilm microalgal system was capable for removing NH4+-N, PO43--P and dissolved COD of 97 %, 93 % and 75 %, respectively at the cultivation time of 14 days, while for suspended microalgal system these were 92 %, 87 % and 68 %, respectively. Biofilm microalgal system also showed advantages of higher biomass production and simple harvesting of biomass, due to it tightly attached on supporting material by the matrix of extracellular polymeric substances (EPS). Moreover, the analysis of potential electricity generation and environmental impact highlighted the promising sustainability of microalgae-based poultry wastes treatment as microalgae provided significant potentials for electricity generation and CO2 reduction. The analysis showed that with nationwide egg-laying hen farms in Thailand, the total electricity generation can be as high as 72 GWh/year with the total CO2 reduction capacity of 99 kton CO2/year, while CO2 emission from electricity generated by microalgal biomass is at least 29 % lower than conventional fuels. The study offers a promising waste management alternative with great potential to achieve efficient treatment and valuable resource recovery for poultry farms in the future.

No abstract available

In this study, the cultivation and harvesting of Arthrospira platensis biomass were proposed via simple, safe, and efficient techniques for direct consumption. Cultivation of microalgae in a covered macrobubble column under outdoor conditions resulted in significant differences (p < 0.05) with a maximum dry cell weight (Xm) of 0.959 ± 0.046 g/L. Notably, outdoor cultures resulted in approximately two-fold biomass compared to indoor cultures. This outcome shows that the developed outdoor setup integrated with solar panels while utilising Malaysia's weather and atmospheric air as carbon sources is viable. Meanwhile, for harvesting, the screening showed that the fungus isolated from mould soybean cake (tempeh) starter indicated the highest harvesting efficiency, which was then further identified as Rhizopus microsporus, microscopically and molecularly. Overall, the economical and portable setup of outdoor cultivation coupled with safe harvesting via locally isolated fungus from tempeh as a bioflocculant would provide sustainability to produce A. platensis biomass.

Background The green alga Chlamydomonas reinhardtii is an accepted food ingredient in the United States of America (United States), the European Union, Singapore, and China. It can be consumed in unlimited quantities. As this alga is rich in nutrients, proteins, and rough polysaccharides and contains a balanced proportion of various amino acids, it is an excellent raw material for food production. Although various edible brown and green algae are available on the market, their color and strong grassy flavor have constrained their popularity among consumers, thereby limiting their application in food additives and animal feed. Methods Chlorophyll-deficient C. reinhardtii mutants were developed using atmospheric and room temperature plasma (ARTP) technology. Results A yellow-colored C. reinhardtii variant (A7S80) cultivated in dark conditions was isolated. This light-sensitive variant has a mutation in the chlM gene, and it can grow heterotrophically using acetate as a carbon source. Conclusion Compared to wild-type C. reinhardtii, A7S80 has significantly lower chlorophyll levels, reduced grassy flavor, and more diverse pigments, with considerable potential for commercial application in human and animal food production, as well as in pharmaceutical and cosmetic industries.

Mushrooms are eukaryotic organisms with absorptive heterotrophic nutrition, capable of feeding on organic matter rich in cellulose and lignocellulose. Since ancient times, they have been considered allies and, in certain cultures, they were seen as magical beings or food of the gods. Of the great variety of edible mushrooms identified worldwide, less than 2% are traded on the market. Although mushrooms have been valued for their multiple nutritional and healing benefits, some cultures perceive them as toxic and do not accept them in their culinary practices. Despite the existing skepticism, several researchers are promoting the potential of edible mushrooms. There are two main methods of mushroom cultivation: solid-state fermentation and submerged fermentation. The former is the most widely used and simplest, since the fungus grows in its natural environment; in the latter, the fungus grows suspended without developing a fruiting body. In addition, submerged fermentation is easily monitored and scalable. Both systems are important and have their limitations. This article discusses the main methods used to increase the performance of submerged fermentation with emphasis on the modes of operation used, types of bioreactors and application of morphological bioengineering of filamentous fungi, and especially the use of intelligent automatic control technologies and the use of non-invasive monitoring in fermentation systems thanks to the development of machine learning (ML), neural networks, and the use of big data, which will allow more accurate decisions to be made in the fermentation of filamentous fungi in submerged environments with improvements in production yields.

Morels are one of the most highly prized edible and medicinal mushrooms worldwide. Therefore, historically, there has been a large international interest in their cultivation. Numerous ecological, physiological, genetic, taxonomic, and mycochemical studies have been previously developed. At the beginning of this century, China finally achieved artificial cultivation and started a high-scale commercial development in 2012. Due to its international interest, its cultivation scale and area expanded rapidly in this country. However, along with the massive industrial scale, a number of challenges, including the maintenance of steady economic profits, arise. In order to contribute to the solution of these challenges, formal research studying selection, species recognition, strain aging, mating type structure, life cycle, nutrient metabolism, growth and development, and multi-omics has recently been boosted. This paper focuses on discussing current morel cultivation technologies, the industrial status of cultivation in China, and the relevance of basic biological research, including, e.g., the study of strain characteristics, species breeding, mating type structure, and microbial interactions. The main challenges related to the morel cultivation industry on a large scale are also analyzed. It is expected that this review will promote a steady global development of the morel industry based on permanent and robust basic scientific knowledge.

Microalgae are fast-growing photosynthetic organisms which have the potential to be exploited as an alternative source of liquid fuels to meet growing global energy demand. The cultivation of microalgae, however, still needs to be improved in order to reduce the cost of the biomass produced. Among the major costs encountered for algal cultivation are the costs for nutrients such as CO2, nitrogen and phosphorous. In this work, therefore, different microalgal strains were cultivated using as nutrient sources three different anaerobic digestates deriving from municipal wastewater, sewage sludge or agro-waste treatment plants. In particular, anaerobic digestates deriving from agro-waste or sewage sludge treatment induced a more than 300% increase in lipid production per volume in Chlorella vulgaris cultures grown in a closed photobioreactor, and a strong increase in carotenoid accumulation in different microalgae species. Conversely, a digestate originating from a pilot scale anaerobic upflow sludge blanket (UASB) was used to increase biomass production when added to an artificial nutrient-supplemented medium. The results herein demonstrate the possibility of improving biomass accumulation or lipid production using different anaerobic digestates.

Edible fungi, as nutritious foods in healthy diets, have gained popularity among consumers. The expansion of the edible fungi cultivation scale led to a shortage of cultivation substrate, making the development and utilization of new substrates a research hotspot. Ginger straw, the main byproduct in the ginger planting process, boasts a huge yield. In this study, ginger straw substrate (GSS) was assessed for the first time for cultivating five major edible fungi. The results indicated a significant improvement in biological efficiency (BE) with GSS, increased by 1.22–64.81%. In terms of nutritional properties, the GSS not only significantly increased the crude protein content (0.36~10.6%) and reduced sugar content (0.01~1%), crude fiber content (0.14~3.87%), and mineral level (The maximum increases were 217.02 mg/kg for calcium, 4.74 mg/kg for magnesium, and 44.08 mg/kg for iron) but also positively affected the total antioxidant capacity and composition of flavor-contributing amino acids. These results provide a scientific basis for cultivating edible fungi with ginger straw and offer a new way for edible fungi substrate selection.

Edible mushroom proteins are gaining recognition as a potentially sustainable alternative to conventional protein sources. However, large-scale development remains limited by extraction inefficiencies and species-dependent cultivation constraints. This review synthesizes current knowledge from a protein-centric perspective, covering nutritional qualities, protein quality indices, bioactive properties of native proteins and derived peptides, and challenges posed by the mushroom cell wall to extraction efficiency. Emerging extraction strategies, including ultrasound-, microwave-, and deep eutectic solvent-assisted approaches, are critically evaluated for their ability to improve protein extraction. These strategies differ in their mechanisms, with ultrasound relying on cavitation-induced physical disruption, microwave extraction enhancing mass transfer through rapid internal heating, and deep eutectic solvents facilitating protein solubilization by modifying cell wall and protein-solvent interactions. Technofunctional properties, particularly emulsification and foaming, point towards encouraging potential, with emerging evidence suggesting that hydrophobins could play a contributing role in these properties. Special attention is also given to the distinctive flavor-binding potential, which may underpin unique sensory attributes in alternative protein systems. Safety considerations, including allergenicity, toxic proteins, and environmental contaminants, are also discussed in relation to their safe utilization. Cultivation constraints and potential sensory issues could limit adoption of mushrooms as an alternative protein source. Overall, in the context of food applications, research in edible mushroom proteins is still at an early stage compared to plant-based proteins and requires more effort to realize its potential.

As the world's population and income levels continue to rise, there is a substantial increase in the demand for meat, which poses significant environmental challenges due to large-scale livestock production. This review explores the potential of microalgae as a sustainable protein source for meat analogues. The nutritional composition, functional properties, and environmental advantages of microalgae are analyzed. Additionally, current obstacles to large-scale microalgal food production are addressed, such as strain development, contamination risks, water usage, and downstream processing. The challenges associated with creating meat-like textures and flavors using techniques like extrusion and emulsion formation with microalgae are also examined. Lastly, considerations related to consumer acceptance, marketing, and regulation are summarized. By focusing on improvements in cultivation, structure, sensory attributes, and affordability, microalgae demonstrate promise as a transformative and eco-friendly protein source to enhance the next generation of meat alternatives.

Current food systems face a paradox: although scientific and technological advances have increased production capacity, they still cannot ensure nutritious and sustainable diets for everyone worldwide. In this context, microalgae stand out as promising bioresources due to their nutritional value, functional properties, and environmental benefits. This review critically examines the current state of microalgae biotechnology for food applications, focusing on cultivation methods, processes, techno-functional properties, regulatory challenges, and consumer perceptions. The analysis indicates that, despite notable progress in cultivation systems and approaches to integration and intensification, high production costs and inconsistent methods of characterizing microalgal biomass remain major obstacles to limit large-scale competitiveness. Additionally, legislation and consumer acceptance issues create a gap between laboratory innovations and industrial implementation. To make microalgae a mainstream ingredient, it is essential: (i) align safety standards and regulations; (ii) incorporate economic feasibility and sustainability; and (iii) develop strategic approaches that translate scientific advancements into practical consumer benefits. Therefore, this study, which explores the intersection of biotechnology, nutrition, and economics, offers a valuable framework to help turn microalgae from a promising idea into a practical solution within global food systems.

Microalgae are promising and sustainable sources of blue food proteins, offering high nutritional quality, environmental resilience, and the potential to meet the rising demand for alternative proteins. Despite these advantages, several challenges hinder their large-scale adoption, including production costs, regulatory barriers, protein extraction difficulties, and consumer perception. This review explores the key factors limiting the use of microalgae in the food industry, addressing economic and technological feasibility, regulatory aspects, and consumer acceptance. The analysis includes commonly used microalgae species, their nutritional profiles, and strategies for optimizing their incorporation into food products. Moreover, developing circular biorefineries and utilizing industrial wastewater for cultivation presents a viable solution to reduce costs and enhance sustainability. Additionally, advancements in protein extraction techniques, combined with technological innovations such as microencapsulation, may overcome sensory challenges, expanding consumer acceptance of microalgae-enriched products. Raising consumer awareness of the nutritional and environmental benefits of microalgae is also crucial for market adoption. Given the global need for sustainable food sources, microalgae represent a viable alternative but require scientific, regulatory, and strategic advancements to become a widely adopted solution in the alternative protein industry.

Aquatic biomass, particularly microalgae and duckweed, presents a promising and sustainable alternative source of plant-based protein and bioactive compounds for food and feed applications. This review highlights the nutritional potential of these aquatic species, focusing on their high protein content, rapid growth rates, and adaptability to nonarable environments. Microalgae, such as Chlorella and Arthrospira spp., and duckweed, such as Lemna minor, are evaluated for their functional food applications, including their roles as protein supplements, bioactive components, antioxidants, and emulsifiers in food formulations. The study also examines their environmental benefits, including wastewater bioremediation, nutrient recycling, and greenhouse gas mitigation, which contribute to a more sustainable agricultural system. Technological advancements in the cultivation, harvesting, and processing of microalgae and duckweed are discussed to enhance their scalability and economic feasibility in food and feed production. The findings suggest that integrating microalgae and duckweed into agricultural and food systems can significantly improve food security, nutritional outcomes, and sustainability. Future research should focus on optimizing cultivation efficiencies, advancing protein extraction techniques, and expanding the applications of aquatic biomass in various food products.

This study presents the first cradle-to-gate life cycle assessment (LCA) of Sargassum naozhouense farming in Xuwen coastal waters, China, following ISO 14040 standards with system boundaries encompassing reproduction, seedling rearing, farming and harvesting in the sea. The findings are discussed within the context of data uncertainty, highlighting avenues for future refinement of life-cycle inventory data for the aquaculture sector. Results demonstrate a carbon footprint of 225.52 kg CO2-eq per tonne of fresh algae, significantly higher than large-scale kelp farming but lower than Porphyra cultivation. Sensitivity analysis identified three prioritized decarbonization pathways: (1) Bamboo consumption (contributing 37.51 % of emissions, primarily for raft frame construction), reducible through optimized rope spacing; (2) 50 % reuse of nylon ropes (critical in seedling cultivation), potentially reducing emissions by 10.97 kg CO2-eq; (3) One-third extension of service life for polypropylene floats, potentially reducing emissions by 3.81 kg CO2-eq. Notably, although cultivation removes 208.63 kg CO2-eq/t of carbon from seawater, this harvested carbon rapidly re-enters the atmosphere through product utilization cycles. As net system emissions exceed carbon removal, current practices fail to constitute an effective carbon sink. Future scale-up should leverage enhanced material reuse and optimized raft design informed by oyster farming, thereby reducing the industry's carbon intensity and environmental impact.

Morchella esculenta Fr., known as Guchi in India, is an edible mushroom from the Ascomycota group. It is nutritious, economically and scientifically valuable. Traditional cultures have long used this mushroom both as food and as a remedy for various health issues. This mushroom is rich in carbohydrates, proteins, fiber, vitamins, minerals, and aromatic compounds. Its unique taste, flavor, and texture make it a popular ingredient in recipes around the world. In addition, Morchella esculenta has several medicinal properties, such as antioxidant, antitumor, antimicrobial, and anti-inflammatory effects, and it is used to aid digestion, act as a body tonic, soothe the skin, and help heal wounds. Research on morel cultivation spans over a century, with China leading the way in large-scale outdoor cultivation. The life cycle of the mushroom involves two main stages: the formation of sclerotia and the production of conidia. Cultivating these mushrooms involves making the spawn, introducing it to a growing medium, adding extra nutrients, managing the fruiting phase, and finally harvesting. The application of exogenous nutrition bags facilitates robust mycelial development. Naturally, Morchella esculenta thrives in cold, hilly regions and is commonly found near hardwood and coniferous trees in a saprobic or mycorrhizal association. Its peak growing season is from March to July, and it is native to the Kullu District in Himachal Pradesh, located in the western Himalayas.

The cultivation of edible mycorrhizal fungi (EMF) has made great progress since the first cultivation of Tuber melanosporum in 1977 but remains in its infancy. Five cultivation steps are required: (1) mycorrhizal synthesis, (2) mycorrhiza development and acclimation, (3) out-planting of mycorrhizal seedlings, (4) onset of fructification, and (5) performing tree orchards. We provide examples of successes and challenges associated with each step, including fruiting of the prestigious chanterelles in Japan recently. We highlight the challenges in establishing performing tree orchards. We report on the monitoring of two orchards established between Lactarius deliciosus (saffron milk cap) and pines in New Zealand. Saffron milk caps yields reached 0.4 and 1100 kg/ha under Pinus radiata and P. sylvestris 6 and 9 y after planting, respectively. Canopy closure began under P. radiata 7 y after planting, followed by a drastic reduction of yields, while P. sylvestris yields still hovered at 690 to 780 kg/ha after 11 y, without canopy closure. The establishment of full-scale field trials to predict yields is crucial to making the cultivation of EMF a reality in tomorrow’s cropping landscape. Sustainable EMF cultivation utilizing trees in non-forested land could contribute to carbon storage, while providing revenue and other ecosystem services.

Microalgae are emerging as promising sources of “blue proteins,” a term used to categorize aquatic proteins derived from marine and freshwater organisms. Producing a large amount of biomass, microalgae are notable for their rich content of amino acids, vitamins, minerals, and bioactive substances. Moreover, advances in cultivation, protein extraction, and product formulation have expanded microalgae's potential as key ingredients in novel and future food systems. Beyond their rich protein content, microalgae can synthesize a wide range of valuable compounds, such as carotenoids, PUFAs, and phycobiliproteins. These substances are known for their antioxidant, anti‐inflammatory, and health‐promoting properties, making them beneficial for use in the food, nutraceutical, and pharmaceutical sectors. Despite these compelling advantages, the large‐scale adoption of microalgae‐based blue proteins remains limited by interconnected challenges. Thus, key barriers include low consumer awareness and limited sensory acceptance, high production and processing costs, lack of standardized regulatory frameworks, and persistent cultural perceptions that associate microalgae with “non‐pleasant” food sources. These issues are complicated by technical hurdles in achieving desirable taste, texture, and color profiles for food applications. Overcoming these constraints will require multidisciplinary efforts that integrate innovations in biotechnology, food processing, sensory science, marketing, and public education. Regarding governments, policy development and investment in infrastructure are also crucial for cost reduction and to scale production sustainably. Therefore, this review presents a comprehensive analysis of key drivers, blockages, and future directions shaping the development of microalgae‐derived blue proteins and their role in building more sustainable, resilient, and nutritious food systems for the future.

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Mycoforestry, a farming system that produces edible fungi crops in forest plantations through controlled mycorrhizal symbiosis, has the potential to enhance biodiversity in forestry plantations and mitigate some of the negative impacts associated with modern agriculture, such as soil erosion, habitat degradation, and carbon emissions. Mycoforestry systems typically exploit a range of native fungi that can be inoculated into planting stock of commercial tree species, with biodiversity benefits delivered through expanded habitat provision for the fungi and a range of other organisms through alterations to stand structure. One mycoforestry system showing strong potential for commercial viability involves the cultivation of Lactarius deliciosus (L.:Fr.) S.F. Gray in Pinaceae plantations. This review aims to evaluate the benefits of mycoforestry systems with a focus on Lactarius deliciosus (L.:Fr.) as a case study. It will review the state of the art and discuss technical developments necessary for the successful large-scale application of mycoforestry systems.

The projected global population of 9.22 billion by 2075 necessitates sustainable food sources that provide health benefits beyond essential nutrition, as the relationship between food biochemistry and human well-being is becoming increasingly significant. Microalgae are simple microscopic organisms rich in various bioactive compounds, such as pigments, vitamins, polyunsaturated fatty acids, polysaccharides, bioactive peptides, and polyphenols, which can be used to develop novel foods with potential health benefits. Bioactive substances offer numerous health benefits, including anti-inflammatory, anticancer, antioxidant, anti-obesity, and heart-protective effects. However, incorporating microalgal biomass into functional food products presents several challenges, including species diversity, fluctuations in biomass production, factors affecting cultivation, suboptimal bioprocessing methods, inconclusive evidence regarding bioavailability and safety, and undesirable flavors and aromas in food formulations. Despite these challenges, significant opportunities exist for the future development of microalgae-derived functional food products. Extensive investigations are essential to overcome these challenges and enable the large-scale commercialization of nutritious microalgae-based food products. This review aims to examine the potential of microalgae as natural ingredients in functional food production, explore the factors limiting their industrial acceptance and utilization, and assess the safety issues associated with human consumption.

Mushroom cultivation represents a crucial sector in sustainable agriculture, offering rich nutrients and economic benefits. However, traditional cultivation methods often lack efficiency and sustainability, compounded by the risk of cultivating inedible or poisonous mushrooms. ML algorithms, such as SMOTE and pipeline models are employed to classify mushrooms into edible or poisonous categories, mitigating the risk associated with cultivation. Agaricus bisporous, or white button mushroom is the most common mushroom grown in many parts of India. The proposed IoT system is designed for convenience and scalability, accommodating both small and large-scale mushroom cultivation operations. The work focuses on efficient and profitable management of mushroom cultivation, providing greater yield and profit.

Abstract The objective of this study was to determine the effects of high pressure to investigate the technical functional properties of the protein solution extracted from an edible insect, Protaetia brevitarsis seulensis. High pressure processing was performed at 0 (control), 100, 200, 300, 400, and 500 MPa at 35°C. The essential amino acid index of the control was lower (p<0.05) than that of the P. brevitarsis seulensis extract treated with 100 MPa. The SDS-PAGE patterns tended to become faint at approximately 75 kDa and thicker at approximately 37 KDa after high pressure treatment. The protein solubility and pH of the protein tended to increase as the hydrostatic pressure levels increased. The instrument color values (redness and yellowness) of the P. brevitarsis seulensis protein treated with high pressure were lower (p<0.05) than those of the control. The forming capacity of the protein solution with P. brevitarsis seulensis treated with high pressure was higher (p<0.05) than that of the control. In conclusion, we confirmed that the technical functional properties of edible insect proteins extracted under high pressure of 200 MPa are improved. Our results indicate that high pressure can improve the technical functional properties of proteins from edible insects.

Insect-based food ingredients are emerging as sustainable protein sources, but their production requires ensuring microbial safety and inactivation of endogenous enzymes to avoid undesirable proteolysis, without compromising protein structure. While traditional thermal processing affects the protein structure, the potential of pulsed electric field (PEF) technology to inactivate microorganisms in lesser mealworm and house cricket slurries at pH 3 while simultaneously retaining the native protein structure is yet unexplored. Lesser mealworm and house cricket slurries at pH 3 were subjected to continuous and batch PEF treatments with varying intensities (0–450 kJ/kg). Microbial inactivation (aerobes, anaerobes, yeasts, and moulds), temperature changes, protein solubility, protein structure (SDS-PAGE and FTIR), and endogenous protease activity were assessed. For both insect species, high-intensity PEF (>150 kJ/kg) achieved up to 5 log microbial reduction, but increased temperatures up to 75 °C, altering protein structure. Low-intensity PEF did not affect protein conformation and protease activity, but was not effective in microbial inactivation (<1 log reduction). We conclude that while PEF can effectively inactivate microorganisms, it cannot be considered a non-thermal method for the present sample conditions due to the temperature increase at higher intensities. PEF could be well-suitable for incorporation in hurdle techniques, such as combinations with moderate heating. Future research should investigate synergistic effects of PEF, also for using alternative PEF set-ups, with other mild processing techniques for effective microbial inactivation while preserving native protein structure. Furthermore, optimal PEF intensities for enhanced protein solubility should be explored.

This study evaluated whether machine learning models could offer improved predictive performance over traditional response surface methodology (RSM) in predicting the solubility of edible insect proteins according to extraction and processing conditions. The machine learning models included linear regression (LR), decision tree (DT), random forest (RF), and XGBoost (XGB). The RSM model yielded statistically significant outcomes (p < 0.001), its applicability across diverse datasets is limited. Meanwhile, DT, RF, and XGB demonstrated exceptional predictive accuracy and reliability, with R2 values greater than 0.99. DT and XGB showed improved metric scores after a 10-fold cross-validation, therefore, compared to RSM, they can be used to predict the unseen data accurately. Additionally, feature importance analysis revealed that the protein extraction methods had a major influence on solubility. These findings suggest that machine learning models can be used to optimize experimental conditions of edible insect proteins research, thus, reducing cost and time.

Simple Summary Many edible insect species are consumed in Africa, but their nutrient composition—taking into account processing methods used to increase their shelf life—is under-documented. To fill knowledge gaps concerning relevant species for cross-border trade throughout Africa, this study analyzed the protein content and the amino acid (AA) profiles of six commercially available species in the Democratic Republic of Congo (DRC). The protein content of the orthopteran and lepidopteran representatives is relatively comparable with values reported for meat, fish, and poultry in the FAO’s food composition database. Some species such as Imbrasia ertli (Lepidoptera) contained high values in essential AAs, supporting the use of edible insects for dietary supplementation for vulnerable populations with cereal-based diets. Furthermore, the study also reported that these insect species could be grouped in three clusters based on their AA profiles, since the AA profiles varied according to insects’ taxa. Representatives of the family Notodontidae contained both the lowest values in several AAs and the essential amino acid index (which is a rapid calculation to determine protein nutritional quality) as compared to Saturniidae and Gryllidae. Overall, our findings supported edible insects as nutrient-rich food and we call for enhancing cross-border trade of species linked to potential economic, social, and ecological benefits. Abstract This study analyzed the protein content of ten edible insect species (using the Dumas method), then focused on the amino acid (AA) profiles of the six major commercially relevant species using HPLC (high-pressure (or performance) liquid chromatography). The protein contents varied significantly from 46.1% to 52.9% (dry matter); the Orthoptera representative yielding both the highest protein content and the highest values in three essential amino acids (EAAs). Regarding Lepidoptera species, the protein content of Saturniidae varied more than for Notodontidae. Imbrasia ertli gave the best example of a species that could be suggested for dietary supplementation of cereal-based diets, as the sample contained the highest values in five EAAs and for the EAA index. Furthermore, first-limiting AAs in the selected insects have also been pointed out (based on a species-specific AA score), supporting that the real benefit from eating insects is correlated to a varied diet. Additionally, preliminary insights into AA distribution patterns according to taxa provided three clusters based on protein quality and should be completed further to help tailor prescriptions of dietary diets. Since the AA composition of the selected insects was close to the FAO/WHO EAA requirement pattern for preschool children and met the requirements of 40% EAAs with high ratio EAAs/NEAAs, the current study endorses reports of edible insects as nutrient-rich and sustainable protein sources.

Chitin present in the shell of edible insects is a potential source of chitin, lipids, and proteins, and it exerts various biological activities. Thus far, only a few studies have focused on the use of chitin as a source of high-protein-diet oligosaccharides. The use of insect chitin for the production of high-protein-diet oligosaccharides can lessen the reliance on diet crops. Moreover, although chitin composition in Tenebrio molitor larva, pupa, and adult has been extensively investigated, chitin extraction from T. molitor larval whole body and exuvium has received poor attention. The present study compared the effectiveness of two techniques for extracting high-protein-diet chitin oligosaccharide from an edible insect (T. molitor). Two different extraction sequences of chitin from the larval stage (molitor stage larvae) and adult stage (molitor stage adult) of edible T. molitor were investigated. Two processing steps were employed: (a) deproteinization (DEP) and (b) demineralization (DEM) treatments. Differences in the order, conditions, and period of their application resulted in two different chitin extraction procedures. The viscosity, degree of polymerization, and crystallinity index of the chitin extracted using the two procedures were measured, and its chemical components (chitin, ash, protein, fat, and moisture contents) were determined. T. molitor adults and larvae treated sequentially with DEM-DEP demonstrated the greatest yield of approximately 14.62 % ± 0.15 and 6.096 % ± 0.10 %, respectively. By contrast, when treated sequentially with DEP-DEM, the recorded yields were 10.96 % ± 0.18 and 5.31 % ± 0.38, respectively. Differences in the degree of deacetylation between both methods were observed. Additionally, Fourier transform infrared spectroscopy and X-ray diffractometry of the extracted chitin along with a commercial sample revealed consistent chain conformation, mean hydrogen bonding, and crystallinity index. In this way, residues produced by farmed edible insects can be recovered and used as a novel source of chitin.

This study explored high-humidity hot air impingement blanching (HHAIB) as a pretreatment to enhance edible crickets' drying efficiency and quality. HHAIB at 120 °C for 3 min reduced drying time by 56 % compared to conventional hot air drying. Scanning electron microscopy showed HHAIB facilitated cuticular wax layer shedding and accelerated dehydration; low-field nuclear magnetic resonance indicated improved water mobility in crickets' tissues post-treatment. After HHAIB at 120 °C for 3 min, the degree of browning and lipid oxidation in crickets was mitigated due to the shortened drying time, with ΔE and Malondialdehyde significantly reduced to 6.13 and 19.15 nmol/g DW. However, the free amino acid and soluble protein contents reduced to 26.26 mg/g DW and 5.71 g/100 g DW due to thermal phenomena. These findings confirm HHAIB as a promising pretreatment for enhancing cricket drying efficiency and quality, providing insights for sustainable insect-food processing development.

As the global population grows, the demand for edible meat is expected to rise. However, traditional livestock farming presents numerous challenges. So alternative proteins emerge as a solution to these issues. This paper analyzes the technological advancements of insect processing and the current status of insect protein market. The processing attaches importance to protein extraction for a superior yield ratio and flavor improvement, which effectively influences consumers’ acceptance. Given that there is no existing regulation or law for insect alternative protein in China, the paper chooses a document from the European Union as a reference. China’s long history of consuming insects provides a favorable environment for the growth of this market. Future development pathways, including the addition of insects to daily recipes, the application of insect-based therapy, and government support, are proposed in this passage. Through these initiatives, insect proteins have the potential to gain wider acceptance and application in the Chinese market.

With the rising demand for sustainable proteins, edible insects such as silkworm pupae are gaining recognition for their high-quality protein and essential nutrients. Advanced technologies like high hydrostatic pressure (HHP) processing have the potential to enhance the functional properties of insect proteins. This study investigated the application of HHP to silkworm pupa protein, focusing on its effects on physicochemical properties, functional characteristics, and bioactivity. HHP treatments at 400 and 600 MPa significantly enhanced emulsifying and foaming capacities, as well as antioxidant activity. Furthermore, HHP-assisted extraction facilitated protein unfolding and increased the exposure of hydrophobic groups on the protein surface, which likely contributed to improvements in protein solubility and antioxidant function. These findings provide valuable insights into the potential of HHP to enhance the quality of edible insect proteins for food applications.

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Processing of edible insects typically involves fractionating into high-value food ingredients, which results in by-products containing chitin and insoluble proteins. This study examined the effectiveness of lactic acid bacteria (LAB) in removing proteins from chitin in insect processing residues. Lesser mealworm processing residues were biologically treated for 48 and 120 h using LAB strains without added carbon sources. Results showed partial deproteinization, up to 29 % with Levilactobacillus brevis after 120 h. Most LAB grew up to 2 log10 colony-forming units/mL in the first 48 h. Confocal microscopy and Fourier-transform infrared spectra indicated that some protein remained attached to chitin. The molecular weight of solubilized proteins was affected by strain and time of incubation, with antioxidant activity increasing significantly after 120 h with Lacticaseibacillus paracasei. The biological treatment of insect processing streams can be a sustainable approach to producing high amounts of LAB biomass with subsequent protein solubilization and chitin release.

Edible insects as an alternative source of protein are gaining increasing attention, leading to new opportunities for their use in food processing. In this study, the functional properties, such as water and oil holding capacity, foaming, and emulsifying properties, of the most popular insect forms (flour, defatted flour, and protein preparations), such as Gryllus asimillis, Acheta domesticus, and Zophobas morio, were studied. Moreover, proximate analysis, protein extraction yield and efficiency, and sensory analysis, were evaluated. Defatting the flours yielded the highest protein content of all the insect forms tested, in the range of 70.51 to 76.02%, significantly reducing their calorific value by up to 35% for Z. morio. Generally, protein preparations exhibit the best functional properties among studied forms, and the most significant differences are noticeable in foaming capacity—near 30% higher than flours. Furthermore, all samples scored well in the sensory test (overall score 3.76–4.47) except for the Z. morio flour (2.93), which may exclude it from being used in the food industry. The results show that the insect forms studied, due to their good functional properties, can become a valuable component of food recipes, positively impacting the characteristics of the designed food.

Advocates for low-fat consumption have shifted focus to developing nutritious low-fat meat products from local, readily available raw materials. Edible African Palm Weevil Larvae (AFPWL) are a widely relished indigenous insect larvae whose properties are affected by processing methods. This study assessed the qualities of sausages produced with differently processed AFPWL. AFPWL, 65-80 days old (n=400), were asphyxiated at 4ºC and processed in three forms: Raw (AFPWL-R), Moist Cooked (AFPWL-MC) and Smoked (AFPWL-S) before use in sausage production. Four types of Sausages (S): AFPWL-RS, AFPWL-MCS, AFPWL-SS and Lard Sausage (LS) were produced. Swelling (%) and Water Absorption Capacities (WAC) (%) were determined on the emulsion. Product yield (PY%), moisture (%), crude protein (%), fat (%), and organoleptic characteristics (9-point hedonic scale) were determined on freshly cooked sausage. Thiobarbituric Acid Reactive Substances (TBARS) (MDAmg/100 g) were assessed over a pooled storage of 21 days. Data were subjected to ANOVA and a significant test using DMRT at Pα0.05. Swelling capacity 9.64 (AFPWL-MCS) and 9.33 (AFPWL-SS) are similar (P >0.05) but significantly higher (P <0.05) than 7.77 (LS) and 6.93 (AFPWL-RS), while WAC 32.00 (AFPWL-SS) is similar to 37.00 (AFPWL-MCS) and 26.00 (AFPWL-RS) but higher (P <0.05) than 23.00 (LS). All AFPWL sausages had higher PY (90.51-96.15) and Moisture contents (61.40-66.59). Crude protein (28.07) AFPWL-MCS and AFPWL-SS (27.91) were higher (P <0.05) than LS (26.53) and AFPWL-RS (24.60). Fat (4.06) in AFPWL-RS was lower (P <0.05) than 4.24 (LS), 4.35 (AFPWL-SS) and 4.46 (AFPWL-MCS). All AFPWL sausages were significantly (P <0.05) tender, but AFPWL-SS sausage was the most acceptable. All AFPWL sausages had significantly higher TBAR values (0.53-0.58) when compared to 0.50 (LS). Processing of AFPPWL before use in sausage production could lead to a novel insect-based meat product with unique, high nutritional and sensory properties. 

Edible insects have emerged as a sustainable alternative protein source, driving recent innovations in the food industry, though food safety and consumer acceptance remain major challenges. This study aimed to develop a quantitative multi-criteria decision analysis (MCDA) method to rank 10 insect-based food products (bar, biscuit, bread, burger patty, fries, milk, nugget, pasta, porridge, and sausage) based on four main criteria (C): consumer intention (C1), environmental impact (C2), nutritional quality (C3), and microbiological food safety risk (C4). Each criterion was defined through a selection of sub-criteria (SC). C1 included daily food intake (SC1.1), consumer proportion (SC1.2), and product preference (SC1.3), with data derived from national dietary surveys and a cross-cultural consumer survey. C2 was assessed through greenhouse gas emissions (SC2.1), land use (SC2.2), water use (SC2.3), and energy resource (SC2.4), using environmental impact database and insect powders processing data. C3 was calculated based on food compositional data and the European Nutri-Score algorithm, while C4 considered the presence likelihood of microbial hazards (SC4.1) and severity of foodborne illness (SC4.2). After normalizing the data and assigning equal weights across criteria and sub-criteria, 10 products were ranked using weighted-sum and ELECTRE III aggregation methods. Sensitivity analyses demonstrated the robustness of ELECTRE III approach. The proposed method provides a practical tool for guiding the development of insect-based foods that are safe, nutritious, sustainable and acceptable to consumers, and can support broader multi-criteria evaluation of novel food products.

As consumer and manufacturer interests in edible insects and processed food with added insects are increasing, new possibilities for detecting edible insect proteins in processed foods have become increasingly important. In the present study, a proteomic strategy was applied to identify insect proteins and thermostable house cricket-specific (Acheta domesticus) peptide markers. To determine the limit of detection (LOD) for house cricket proteins, cooked meatballs containing house cricket protein powder (CP) as a partial pork substitute were investigated. The final concentration of CP ranged from 0.8% to 7.6%. The LODs for tropomyosin 1 and translational elongation factor-2 were 0.8% (w/w), whereas for apolipophorin-III it was 2.5% (w/w). Eight heat-resistant peptides unique to the family Grillidae (true crickets) and four peptides unique to the Acheta domesticus were identified. The results suggest that selected cricket-specific and processing-resistant peptide markers have potential utility in the authentication of the cricket formulations used in meat products. However, this has to be confirmed on different heavily processed meat products.

The potential of edible insects in alleviating the nutritional challenges facing humans has led to an increased interest in their utilization.  Edible insects contain substantial amounts of protein and fats, which are very important macronutrients that confer useful functional and physicochemical properties to foods.  However, the functional properties of these nutrients have not been given sufficient attention, limiting their use as ingredients in diets and food products.  Processing of insects converts them into an ingredient that can be added to various foods, improving their acceptability in terms of flavor, taste, and nutritional content.  Popular processing methods for edible insects include frying, boiling, oven drying, roasting, smoking, and toasting.  These thermal treatments reduce the insect water content, enabling them to be milled into powders.  This study set out to investigate the fatty acid profile, protein digestibility, and physicochemical and functional properties of powders and oils of edible dung beetle (Scarabaeus satyrus).  The dung larvae were collected from three counties in Western Kenya.  They were cleaned and subjected to drying and milling, then analyzed.  Results indicated that protein digestibility ranged between 64.27-70.03%.  The dominant unsaturated fatty acid was Oleic (45.71±2.45%), while the main saturated fatty acid was Lauric (45.43 ±0.8%).  The saponification value of the oils ranged between 127.94 -130.17 mgKOH/g oil, Acid value (41.96-44.11 mg KOH/g oil), peroxide value (4.23-3.8 Meq.thio / kg sample), refractive index (1.41-1.44/25ºC) and Iodine value (77.89-88.02 g I/100g).  Functional properties of the powders showed high lipophilic (332.45±19.73%) and moderate (1.11±0.14 ml/g) hygroscopic tendencies in toasted samples from Bungoma and Siaya Counties, respectively.  Emulsifying capacity varied between 80.85%-82.53%, while emulsifying stability ranged from 80.85%-81.33%.  These findings show that the edible Scarabaeus satyrus can provide unsaturated fatty acids and can be a good ingredient in foods as an alternative to conventional cuisines

Entomophagy (insect eating) is an age long practice by man all over the world. The nutritive values of insects are affected by many factors including processing methods. Grasshopper is one of the edible insects in Nigeria. This study aimed at evaluating the effects of processing methods on the nutritive value of adult variegated grasshopper, Zonocerus variegatus. The grasshoppers were collected from an uncultivated farmland, washed, degutted and processed in three ways namely; boiling, roasting and frying. Processed Z. variegatus were analyzed for their proximate, minerals, vitamins and anti-nutrient profile. The data obtained were analyzed by One Way Analysis of variance and means separation was done by Duncan Multiple Range Test. The roasted grasshoppers were found to have the highest value of protein (41.54%) while the boiled insects recorded the least value (18.12%). The roasted also had the highest crude fibre (8.38%), while the raw had the least value (5.04%). The highest crude fat (37.11%) was found in the fried grasshoppers while the boiled had the least crude fat (1.59%). The fried grasshoppers recorded highest value of carbohydrate (3.43%). The roasted insects had the highest values in all the eleven minerals analyzed. Lead and Nickel were not detected in all the samples. Potassium was the highest across all processing methods, ranging from 88.01- 465.99 mg/100g while manganese was the least (0.44-1.46 mg/100g). Vitamins A, E, B2, B6 and B12 were also examined. The values for all these were highest in the roasted with 0.45, 0.48, 0.013, 0.99, and 2.63 respectively. The boiled grasshoppers had the least values in all the parameter examined except in Vitamin B6 where it had a higher value of 0.84) than the fried (0.79) and raw (0.67). Tannin was highest in the fried grasshopper (4.51) and the least in raw grasshoppers (1.29). It can therefore be concluded that processing methods significantly affected the nutritive value of Z. variegatus.

Understanding the impacts of protein production systems is necessary to plan the just transition of food systems. We analysed 285 studies to assess the impacts of 13 protein systems across 25 indicators under five key categories—natural capital, human capital, social capital, produced capital and governance. Nine protein systems (regenerative, organic, rangelands, free-range poultry, sustainable energy cultivated meat, conventional energy cultivated meat, mixed grains and livestock, pastoralists and plant-based) have overall positive impacts across all five categories. In comparison, four protein systems have negative impacts (small-scale beef, caged poultry, industrial pork, and confined feeding operations). We then used this in-depth assessment to develop five ‘what if’ future scenarios to track and assess the transition of protein production systems to 2050. Rapid reduction of industrial production may contribute to a just and inclusive transition of protein production systems. This assessment can help reduce risks associated with negative impacts and assist in governing and managing protein production systems towards long-term sustainability.

The possibility of using complex plant raw materials for microbial biomass production has been investigated. The chemical composition of different types of plant raw materials (grasses, beetroot, potatoes, corn, waste) of the northeastern region was analysed, which made it possible to develop an optimal nutrient medium for the cultivation of Candida scotti Tul-1. Cultivation was carried out in flasks and fermenter, controlling temperature, pH and aeration. The optimum concentration of sugar in the medium was found to be 4% and the culturing temperature was 28°C. The obtained biomass contained 60-62% protein with a balanced amino acid composition. The results demonstrate the promising use of plant raw materials for microbial protein production and waste utilisation. The developed technology can become the basis for sustainable and economically favourable production of protein feeds and food supplements. Further research is aimed at optimising the process and scaling up production.

No abstract available

Digital transformation is driving more precise, efficient, and sustainable practices in both farming and postharvest crop processing. As consumer preferences shift toward healthier alternative protein sources, pulses are gaining attention with their market currently valued at USD 78 billion. However, challenges related to cultivation, protein quality, and extraction efficiency limit their full potential in both food and non‐food applications. This review investigates emerging technologies aimed at enhancing pulse crop cultivation and protein fractionation processes. Tools such as artificial neural networks (ANNs), geospatial analysis, and remote sensing are explored for their ability to optimize pulse crop production by improving yield prediction, protein content assessment, and environmental adaptation. Additionally, an overview of dry and wet protein fractionation techniques is discussed, with an emphasis on optimizing processes through meta‐analysis, response surface methodology (RSM), and discrete element method (DEM). These methods enhance the efficiency of these fractionation methods, while boosting protein purity, functionality, and digestibility. The integration of advanced analytical and computational strategies in precision agriculture and pulse processing can help overcome current limitations and support the expanding pulse protein market.

The increasing global demand for sustainable protein sources necessitates the exploration of alternative solutions beyond traditional livestock and crop-based proteins. Microalgae present a promising alternative due to their high protein content, rapid biomass accumulation, and minimal land and water requirements. Furthermore, their ability to thrive on non-arable land and in wastewater systems enhances their sustainability and resource efficiency. Despite these advantages, scalability and economical feasibility remain major challenges in microalgal protein production. This review explores recent advancements in microalgal protein cultivation and extraction technologies, including pulsed electric field, ultrasound-assisted extraction, enzyme-assisted extraction, and microwave-assisted extraction. These innovative techniques have significantly improved protein extraction efficiency, purity, and sustainability, while addressing cell wall disruption and protein recovery challenges. Additionally, the review examines protein digestibility and bioavailability, particularly in the context of human nutrition and aquafeed applications. A critical analysis of life cycle assessment studies highlights the environmental footprint and economical feasibility of microalgal protein production compared to conventional protein sources. Although microalgal protein production requires significant energy inputs, advancements in biorefinery approaches, carbon dioxide sequestration, and industrial integration can help mitigate these limitations. Finally, this review outlines key challenges and future research directions, emphasizing the need for cost reduction strategies, genetic engineering for enhanced yields, and industrial-scale process optimization. By integrating innovative extraction techniques with biorefinery models, microalgal proteins hold immense potential as a sustainable, high-quality protein source for food, feed, and nutraceutical applications.

The environmental burden of widely used protein sources in animal feeds, such as soybean and fishmeal, has raised concerns about the sustainability of current livestock production systems. In response, alternative protein sources are being explored, with insect meal emerging as a promising candidate. This study conducted a comparative Life Cycle Assessment (LCA) of four compound pig feed formulations differing in protein composition, incorporating soybean meal, fishmeal, and Tenebrio molitor (insect) meal. The LCA followed ISO 14040/44 standards and applied both mass-based and protein-based functional units (FUs) to examine how FU choice influences environmental outcomes. Results showed that crop-derived ingredients, particularly soybean meal, drove most environmental burdens due to land use change and fertilizer inputs. Replacing soybean with insect meal led to impact reductions in key environmental categories. Insect meal’s scalability, efficient land use, and potential waste valorisation supported its role as a sustainable alternative. The study also highlighted key sustainability issues not assessed by LCA, such as overfishing and ecosystem disruption, associated with fishmeal. Overall, insect meal appeared to be a strong replacement for soybean and fishmeal, with soy substitution proving key to reducing environmental burdens. Finally, the protein-based FU was more relevant given the study’s nutritional focus.

The growing demand for sustainable, nutritionally adequate plant-based foods has driven innovation in meat analogues. This study presents a novel approach to upcycling potato juice protein—a by-product of starch production—into plant-based gyros (PBG) enriched with iron and dietary fiber. Four formulations (PBG1–PBG4) were developed using a blend of potato, rice, wheat, and pea proteins, and fortified with either ferritin-rich sprout powder or ferrous sulfate. Comprehensive analyses were conducted to assess nutritional composition, mineral content, glycoalkaloid safety, antioxidant activity, texture, water mobility, sensory appeal, and microbiological stability. All variants met high-protein labeling criteria and exhibited favorable fiber and mineral profiles. In vitro digestion significantly enhanced antioxidant bioaccessibility, particularly phenolic acids. Sensory evaluations favored ferritin-enriched variants, which also demonstrated superior texture and consumer acceptance. Microbiological assessments confirmed safety for up to 10 days under refrigeration. These findings highlight the potential of potato juice protein as a sustainable, functional ingredient in next-generation plant-based meat analogues.

Accelerating emergence of antimicrobial resistance among food pathogens and consumers’ increasing demands for preservative-free foods are two contemporary challenging aspects within the food industry. Antimicrobial packaging and the use of natural preservatives are promising solutions. In the present study, we used beta-casein—one of the primary self-assembly proteins in milk with a high polymeric film production capability—as a fusion partner for the recombinant expression of E 50-52 antimicrobial peptide in Escherichia coli. The pET21a-BCN-E 50-52 construct was transformed to E. coli BL21 (DE3), and protein expression was induced under optimized conditions. Purified protein obtained from nickel affinity chromatography was refolded under optimized dialysis circumstances and concentrated to 1600 µg/mL fusion protein by ultrafiltration. Antimicrobial activities of recombinant BCN-E 50-52 performed against Escherichia coli, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus aureus, Aspergillus flavus, and Candida albicans. Subsequently, the synergistic effects of BCN-E 50-52 and thymol were assayed. Results of checkerboard tests showed strong synergistic activity between two compounds. Time–kill and growth kinetic studies indicated a sharp reduction of cell viability during the first period of exposure, and SEM (scanning electron microscope) results validated the severe destructive effects of BCN E 50-52 and thymol in combination on bacterial cells.

Purpose Recently, demand for plant-based milk products (PBMP) has increased for multiple reasons, such as the rapid population growth expected to reach 9.7 billion by 2050, health concerns such as lactose intolerance, nutritional aspects, ethical reasons, and environmental concerns. This leads to increased demand for food and competition for natural resources. Hemp-based milk is an emerging dairy alternative, and stakeholders in the supply chain are becoming increasingly interested in learning about the environmental effects of its production. This article aims for a comparative life cycle assessment of hemp-based and bovine milk with fat and protein correction to account for the differences in macronutrient content. Methods The cradle-to-factory gate LCA relied on experimental cultivation and milk production in Lower Saxony, Germany. Inventory was based on primary data from fields and the pilot plant of DIL e. V. and on literature and ecoinvent database to develop a life cycle assessment (LCA) model. The LCA was performed using Simapro 9.3 software and IMPACT 2002+ impact assessment method. The life cycle stages include cultivation, harvesting, and milk production. The study compared hemp-based milk to bovine milk based on 1 kg fat and protein-corrected milk (FPCM) as a functional unit (FU). Co-products are taken into consideration using mass-economic allocation. Results The results showed that hemp cultivation accounted for the highest impact (99%) in the production chain of hemp milk production. The GWP of 1 kg of FPCM hemp-based milk is 0.42 kg CO_2 eq. The energy consumption for 1 kg of FPCM hemp-based milk is 4.73 MJ (12.26% lower than bovine milk). The other main factors impacting hemp-based milk production were terrestrial ecotoxicity (6.444E2 kg TEG soil) and aquatic ecotoxicity (2.458E2 kg TEG water). Hemp fiber was the co-product with 40% of the allocated impacts. The results are sensitive to the changes in fat-protein contents, functional unit, and system boundaries. The results demonstrated that the impacts of hemp milk production were within the range indicated for other PBMP production and 51.7% lower than bovine milk production in terms of GWP. This range primarily stems from field emissions, fertilizer application, and machinery usage during cultivation and harvest. Conclusion The results of the comparisons of bovine milk and hemp-based milk were dependable on the FU. The hemp-based milk has the potential to be a more sustainable alternative to bovine milk due to considerably lower impacts in impact categories—land occupation (99% lower than bovine milk), global warming (52% lower than bovine milk), and ionizing radiation (23% lower than bovine milk). It is primarily due to less use of agricultural machinery, less land requirement, and lower NH_3 emissions than bovine milk in various stages of milk production.

Aim: To evaluate the physicochemical, microbial, sensory attributes and storage stability of coconut yoghurt fortified with date syrup. Methodology: Yoghurt was prepared from coconut milk and date syrup. Five samples were formulated as follows: T0 (100% cow milk), T1 (100% coconut milk), T2 (85% coconut milk, 15% date syrup), T3 (80% coconut milk, 20% date syrup), T4 (75% coconut milk, 25% date syrup) and T5 (70% coconut milk, 30% date syrup) The yoghurt samples were subjected to physicochemical and microbial analysis using standard methods. The sensory attributes assessed were appearance, aroma, taste, consistency and overall acceptability. The pH, titratable acidity and viscosity were evaluated during 4 weeks of storage at refrigerated condition (4- 6o c). Results: From the result, pH, titratable acidity, viscosity, and total solid ranged from 4.50 - 4.41%, 0.82 – 0.94%, 12.0 -72.40 cP and 11.64 – 18.81% respectively. These parameters increased significantly (p<0.05) with increase in the proportion of date syrup. The result showed a significant increase in ash, protein, carbohydrate, and fiber with a corresponding decrease in fat and moisture content as the proportion of date syrup increased. The total bacterial count ranged from 6.95 - 8.20cfu/ml, with Yeast, molds and coliforms undetected. The produced yoghurt samples all have quality attributes of animal milk yoghurt and better storage stability. Conclusion: Therefore, production of commercial yoghurt using plant-based milk (coconut milk) blended with nutritiously rich sweeteners like date fruit is encouraged.

Significance The cultivation of microbial biomass, which is rich in proteins as well as other nutrients, can play a vital role in achieving food security while mitigating the negative environmental footprint of agriculture. Here, we analyze the efficiency associated with using solar energy for converting atmospheric CO2 derived from direct air capture into microbial biomass that can feed humans and animals. We show that the production of microbial foods outperforms agricultural cultivation of staple crops in terms of caloric and protein yields per land area at all relevant solar irradiance levels. These results suggest that microbial foods could substantially contribute to feeding a growing population and can assist in allocating future limited land resources. Population growth and changes in dietary patterns place an ever-growing pressure on the environment. Feeding the world within sustainable boundaries therefore requires revolutionizing the way we harness natural resources. Microbial biomass can be cultivated to yield protein-rich feed and food supplements, collectively termed single-cell protein (SCP). Yet, we still lack a quantitative comparison between traditional agriculture and photovoltaic-driven SCP systems in terms of land use and energetic efficiency. Here, we analyze the energetic efficiency of harnessing solar energy to produce SCP from air and water. Our model includes photovoltaic electricity generation, direct air capture of carbon dioxide, electrosynthesis of an electron donor and/or carbon source for microbial growth (hydrogen, formate, or methanol), microbial cultivation, and the processing of biomass and proteins. We show that, per unit of land, SCP production can reach an over 10-fold higher protein yield and at least twice the caloric yield compared with any staple crop. Altogether, this quantitative analysis offers an assessment of the future potential of photovoltaic-driven microbial foods to supplement conventional agricultural production and support resource-efficient protein supply on a global scale.

Cultivating meat is a promising solution to the negative problems brought by traditional animal husbandry. To make cultured meat have the sensory and nutritional characteristics of conventional meat as much as possible, many studies have been conducted on various cell types and scaffold characteristics. Therefore, this study aims to produce a low-cost cultured meat with a quality closer to that of conventional meat. Tissue generation requires three-dimensional (3D) scaffolds to support cells and simulate extracellular matrix (ECM). Here, we used peanut wire-drawing protein (a biomaterial based on edible porous protein) as a new culture meat scaffold to culture cells. The scaffold can support cell attachment and proliferation to create 3D engineered porcine muscle tissue. The differentiation of smooth muscle cells (SMCs) was induced by a low serum medium to produce more extracellular matrix proteins. After differentiation, it was found that peanut wire-drawing protein scaffolds could be used for porcine smooth muscle cell adhesion and growth. The ECM protein and muscle protein produced by SMCs can endow cultured meat with better quality. This technology provides an innovative pathway for the industrialized production of cultured meat.

No abstract available

Background: Global food security faces significant challenges, particularly with projected rapid population growth and limited natural resources. In an effort to support sustainable food security, this study aims to develop hybrid protein cookies that combine animal and plant proteins and implement a two-phase natural preservation system. Previous studies have shown that the use of hybrid proteins can improve the nutritional quality of food products, while natural preservation systems can reduce dependence on harmful synthetic preservatives. Methods: This study used a comprehensive literature review method to formulate optimal formulation, as well as experiments to evaluate the effectiveness of a combination of whey protein, pumpkin seed flour, and mung bean in improving the nutritional profile and shelf life of cookies. Findings: The results showed that these hybrid protein cookies have a higher protein content, a softer texture, and a shelf life up to 40% longer compared to conventional cookies. Conclusion: These findings support the importance of utilizing dairy industry waste such as whey protein, as well as local resources in improving sustainability and food security. In conclusion, this hybrid protein cookie innovation offers a new solution in creating nutritious, environmentally friendly food products and supporting local economic empowerment. Novelty/Originality of this article: The original aspect of this research is the application of a natural preservation system using mangosteen peel extract, which provides a new contribution to the development of sustainable functional food products.

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In the context of dietary transition, blending animal-source protein with plant-source protein offers a promising way to exploit their nutritional complementarity. This study investigates the feasibility of formulating an iron-rich hybrid food product blending plant-source and animal-source protein ingredients for iron-deficient populations. Using a commercial 3D-food printer, two different-shaped products composed mainly of pork and chicken liver and red lentils were designed. After baking at 180 °C with 70% steam, the 3D-printed products were packed under two different modified atmospheres (MAP): O2-MAP (70% oxygen + 30% carbon dioxide) and N2-MAP (70% nitrogen + 30% carbon dioxide) and stored at 4 °C. pH, water content, aw, lipid oxidation, heme iron and non-heme iron contents and textural properties were measured after 0, 7, 14 and 21 days in storage. After 21 days in storage, the 3D-printed hybrid products had an iron content of around 13 mg/100 g, regardless of the product form and packaging method. However, O2-MAP products showed significant (p < 0.05) time–course changes from day 0 to day 7, i.e., an increase in lipid oxidation, a decrease in heme iron content and an increase in product hardness, gumminess and chewiness. This work opens prospects for developing hybrid food products that upvalue animal by-products.

Searching for natural scaffolds with structural and nutritional properties favorable for cell adhesion and growth is a challenge for piscine cell culture. In this study, myoblasts and preadipocytes from Larimichthys crocea were cultured on uncoated rice grains, and the cell confluence reached over 80% after two days of cultivation. The physical and chemical properties of different rice grains showed that the hardness had a relatively significant regulatory effect on the growth of cells, and 6.45 to 7.28 N was beneficial for the growth of cells. Finally, the nutritional composition and flavor of rice grains were evaluated, revealing that myotubes-organized rice grains were rich in protein, adipocytes-organized rice grains were high in fat. All rice grain samples displayed enhanced flavor profiles. These findings suggest that specific rice varieties can serve as effective scaffolds for the in vitro proliferation and differentiation of piscine stem cells. The resulting rice-meat composite food emerges as a promising innovative food product.

Cultured meat technology is a promising new technology to solve the negative problems brought by traditional animal husbandry. Cultured meat should be further developed to appear on consumers' tables as alternative protein product. Therefore, this study used food grade peanut wire-drawing protein as scaffold to culture smooth muscle cells (SMCs) in vitro to obtain cultured meat productions containing both animal protein and plant protein. Multiple passages lead to the decline of the proliferation rate of SMCs in the proliferation stage and the differentiation ability in the differentiation stage, which means that the plasticity of cells decreased in the later stage of passage. SMCs can well adhere to the peanut wire-drawing protein scaffold and produce extracellular matrix protein and muscle protein, so as to form a cultured meat product with rich protein composition. This study provides a theoretical basis for the production of nutrient-rich cultured meat products.

Meat analogues have been gaining popularity as a sustainable and health-conscious alternative to traditional meat products, driven by increasing consumer awareness of environmental benefits. However, there remains a gap in the market for meat analogues that not only mimic meat texture but also offer enhanced nutritional benefits, particularly in terms of fiber content. Meat analogue strips from Ulva ohnoi and surimi underwent proximate, fiber, and texture profile analysis. The aim of the study was to determine the best formula between the ratio of Ulva ohnoi and surimi inclusion in meat analogue strips along with evaluating the physicochemical characteristics of the resulting product. This study also involved panelists to assess the hedonic level of consumer acceptance of the produced products. The results showed that F5, with 5% Ulva ohnoi and 20% surimi, was the best formula with a sensory value of color 7.21 and texture 6.08. The chemical composition of meat analogue strip F5 includes 2.63 ± 0.54% moisture content, 10.12 ± 2.28% ash content, 3.89 ± 0.87% fat, 13.86 ± 0.31% protein, 59.26 ± 1.93% carbohydrate, and 10.23 ± 0.00% fiber. Eventually, this study opens up new alternatives, showing that meat analogue strips not only serve as an energy source but also as a potential high-fiber healthy snack.

It is important to have sustainable and edible scaffolds to produce cultivated meat. In this research, three-dimensional (3D) porous scaffolds were developed by soy protein amyloid fibrils for cultivated meat applications. Food-safe biological and physical cross-linking methods using microbial transglutaminase and temperature-controlled water vapor annealing technique were employed to crosslink soy protein amyloid fibrils, resulting in the production of 3D scaffolds. The generated 3D scaffolds had pores with sizes ranging from 50 to 250 μm, porosities of 72-83%, and compressive moduli of 3.8-4.2 kPa, depending on the type of soy protein used in the process (β-conglycinin (7S), glycinin (11S) and soy protein isolate (SPI)). When present with pepsin, these scaffolds can degrade within an hour but remain stable in phosphate-buffered saline for at least 30 days. The soy protein amyloid fibril scaffolds enabled C2C12 mouse skeletal myoblasts proliferate and differentiate without adding cell adhesive proteins or other coatings. The results demonstrate the potential of abundant and inexpensive soy protein amyloid fibrils to be utilized as scaffold materials for cultivated meat in the food industry.

A novel hybrid protein gel was developed to sustainably meet the growing demands for protein with pea and whey protein isolates (3:2 in 15 % w/v protein content) which was ultrasound treated (7.5 and 15 min), and gelled (90 °C, 60 min). The study investigated the impact of ultrasound treatment on the structure and gastric breakdown kinetics of hybrid protein gels and compared these properties to commercial protein-based foods (ham, paneer, and mozzarella). Ultrasound treatment for 15 min significantly (p < 0.05) reduced particle size (d50: 5.4 µm vs 32.5 µm in control) and resulted in a higher initial Young's modulus than control. Protein hydrolysis at 180 min was ∼53 % lower (p < 0.05) with 15 min ultrasound treatment than control and 7.5 min ultrasound treatment. Hybrid protein gels exhibited similarities in initial Young's modulus to mozzarella (p > 0.05), while ham and paneer were significantly firmer (p < 0.05). Effective diffusivity of moisture from gastric fluid decreased (p < 0.05) in the order: ham > paneer and mozzarella > hybrid protein gels. In contrast, the effective acid diffusivity from gastric fluid was similar (p > 0.05) between hybrid protein gels and paneer, which were ∼74 % higher (p < 0.05) than ham and mozzarella. Digestion time influenced (p < 0.05) breakdown mechanisms (texture, dry matter loss, moisture, and acid uptake) during digestion. This study confirmed that hybrid protein gels were comparable to commercial protein-based foods and the limiting factor driving gastric breakdown is unique to different foods incorporating proteins.

Hybrid meat products, involving the partial replacement of meat with plant proteins, provide a promising solution to address the rising consumer demand for sustainable and healthier food options. Rice protein (RP), featuring balanced amino acid composition and hypoallergenic property, represents a potent alternative to common legume-based proteins. This study investigates the potential of RP as a partial replacer for pork in kung-wan, a traditional Oriental-style emulsified meatball. RP was incorporated at substitution levels of 10 %, 20 %, and 30 %, and its influence on physicochemical properties, microstructures, and sensory attributes were evaluated. Increasing RP proportion led to reduced emulsion stability, cooking yield, and texture, with significant changes observed at levels above 20 %. Additionally, microstructural analysis revealed that RP substitution caused fat globule agglomeration and transformed the protein matrix from fine-stranded filaments into amorphous aggregates. Moreover, sensory evaluation indicated that a 10 % substitution preserved consumer preference, whereas higher levels caused noticeable declines in hardness and overall acceptability. Furthermore, transglutaminase (TGase) treatment enhanced the cohesiveness of the protein network in RP-hybrid kung-wan at the 20 % substitution level, restoring the texture attributes and sensory qualities to levels comparable to those of the full-meat counterpart. This study highlights the feasibility of RP in hybrid meat products and unveils the role of TGase in facilitating higher substitution levels without compromising products quality.

Strategic regulation of protein coaggregation via molecular interactions enables sustainable production of gel-based foods with reduced economic and ecological effects. This study explored the synergistic effect of soy glycinin (11S) on the heat-induced aggregation of cod proteins (CPs), focusing on aggregate formation pathways and molecular interactions. Incorporation of 11S restructured the secondary structure of CPs with increased β-sheet content, which facilitated coaggregation and produced large-sized complexes with low molecular weight. At 100 °C, the dissociated basic subunit of 11S preferentially bound to myosin heavy chains and actin, triggering further assembly through hydrophobic interactions and disulfide linkages, with hydrophobic forces being dominant. Moreover, acidic pH and salt ions contributed to synergistic enhancement in the coaggregation behavior, with Ca2+ inducing additional cross-linking between proteins via actin-binding sites. These findings highlight 11S as a native, sustainable cross-linker to enhance CP aggregation, fostering the engineering advances of clean-label hybrid protein foods.

The partial or total replacement of proteins from animal sources by those from plant sources is a useful strategy to increase sustainability aspects of diets. Thus, the production of hybrid protein systems is relevant in this context of food transition. Aiming to increase the industrial relevance of corn gluten meal (CGM), a co-product of corn starch production, microfluidization was used and CGM colloidal and emulsifying properties were evaluated alone and in combination with whey protein concentrate (WPC). Microfluidization significantly improved the dispersibility of CGM (reduction of D90 in around 97 %). Dispersions (2 % w/ w, pH 7.0) of microfluidized CGM, WPC and a balanced mixture of these two proteins (50:50) were studied before and after heat treatment (90 °C per 15 min). Before heat treatment, the emulsifying properties of WPC were significantly (p < 0.05) better than those of CGM, for all evaluated parameters. Under these circumstances, the emulsifying properties of the hybrid system containing the two protein sources were dictated by the WPC. After heat treatment, the aggregation of the whey proteins led to a significant reduction in the emulsifying properties of the WPC. Consequently, CGM proteins began to coexist with whey proteins at the oil-water interface of the hybrid emulsions. Despite the overall superior emulsifying properties of WPC, the results suggest that microfluidized CGM may contribute to the development of hybrid protein systems with WPC for applications involving severe heat treatments.

There is a need to reduce the proportion of animal-derived food products in the human diet for sustainability and environmental reasons. However, it is also important that a transition away from animal-derived foods does not lead to any adverse nutritional effects. In this study, the potential of blending whey protein isolate (WPI) with either shiitake mushroom (SM) or oyster mushroom (OM) to create hybrid foods with enhanced nutritional and physicochemical properties was investigated. The impact of OM or SM addition on the formation, microstructure, and physicochemical attributes of heat-set whey protein gels was therefore examined. The mushroom powders were used because they have relatively high levels of vitamins, minerals, phytochemicals, and dietary fibers, which may provide nutritional benefits, whereas the WPI was used to provide protein and good thermal gelation properties. A variety of analytical methods were used to characterize the structural and physicochemical properties of the WPI-mushroom hybrids, including confocal microscopy, particle electrophoresis, light scattering, proximate analysis, differential scanning calorimetry, thermogravimetric analysis, dynamic shear rheology, textural profile analysis, and colorimetry. The charge on whey proteins and mushroom particles went from positive to negative when the pH was raised from 3 to 9, but whey protein had a higher isoelectric point and charge magnitude. OM slightly increased the thermal stability of WPI, but SM had little effect. Both mushroom types decreased the lightness and increased the brownness of the whey protein gels. The addition of the mushroom powders also decreased the hardness and Young’s modulus of the whey protein gels, which may be because the mushroom particles acted as soft fillers. This study provides valuable insights into the formation of hybrid whey protein-mushroom products that have desirable physiochemical and nutritional attributes.

Given the rising demand for more sustainable, cookable dairy alternatives, this research explores the formation and characteristics of heat- and acid-induced gels combining micellar casein and pea protein. Protein dispersions (4 % w/w) of commercial micellar casein isolate and pea protein isolate were prepared and preheated (95°C, 30 min) separately before mixing in varying ratios (75:25 %, 50:50 %, and 25:75 % w/w). After emulsifying with milk fat (3.5 % w/w), the protein mixtures were heated to 80 °C and acidified to pH 5.2 (citric acid). The resultant coagula were pressed, drained, and molded to obtain the final gel. It was observed that adding pea protein led to a higher yield of coagula with more serum retained. As the proportion of pea protein increased, the total solids (TS), protein, and fat content of the gels decreased linearly. The micellar casein gel showed significantly higher hardness, elasticity, and chewiness than the gels containing pea protein. Moreover, the micellar casein gel did not show clear fracture behavior under large deformation, while the gels containing pea protein were more prone to rupture. These textural differences were explained by the changes in gel compositions, protein interactions, and gel microstructure. The composition and textural properties of hybrid gels showed a strong linear relationship with pea protein fractions, showing the possibility of customizing gel properties. Notably, the hybrid gel containing 25 % pea protein exhibited promising characteristics, closely resembling those of the commercial dairy paneer product.

Milk proteins are known for their exceptional nutritional and technological attributes, making them a staple in the food industry. Nonetheless, the partial substitution of milk proteins with plant‐based proteins in dairy products may be an effective strategy to meet the increasing consumer demand for a reduction in the consumption of animal‐derived proteins. This study aimed to evaluate the impact of partial substitution (25% and 50%) of milk proteins with pea protein on the manufacturing and technological attributes of high‐protein yogurt during refrigerated storage. The replacement of up to 50% of the milk proteins with pea protein did not alter the fermentation time and all yogurts had a total lactic acid bacteria count greater than 107 CFU g−1 after manufacturing. However, replacing 50% of milk proteins with pea protein affected the pH, syneresis, water holding capacity, consistency, firmness, viscosity index, and cohesiveness of the yogurts. In turn, no effect was observed on the pH, syneresis, water holding capacity, firmness, and cohesiveness of the product after replacing 25% of the dairy base with pea protein (P >0.05). Hence, the findings indicate that substituting 25% of the milk protein with pea protein in high‐protein yogurts can be achieved without compromising the product's stability.

BACKGROUND Plant-derived proteins are rapidly emerging as innovative ingredients in the food sector because of their sustainability and ethical benefits compared with animal-based proteins. Among dairy applications, fermented beverages are the most suitable products for the incorporation of these proteins. This study evaluated how cold plasma (CP) treatment time and concentrations of modified pea protein isolate (PPI) affected the quality and stability of a hybrid dairy beverage. RESULTS Higher PPI levels increased titratable acidity, whereas CP-treated PPI resulted in higher pH values reaching 4.36 at 1.8 g 100 mL-1. Both PPI concentration and CP treatment improved the water-holding capacity (WHC), with a maximum WHC of 29.68% achieved at 1.8 g 100 mL-1 PPI and a CP treatment time of 30 s. Longer CP time and higher PPI levels increased a* and b* values significantly (P < 0.05). Viscosity peaked at 214.35 Pa s at 1.8 g 100 mL-1 PPI and a CP treatment time of 60 s but declined at 120 s. Pea protein isolate also promoted Streptococcus and Lactobacillus spp. growth, especially with shorter CP time (P < 0.05). CONCLUSION These findings suggest that brief CP treatment is an effective approach to modifying PPI, improving its suitability for use in the development of functional hybrid fermented dairy beverages. © 2026 Society of Chemical Industry.

The escalating global protein demand and the environmental burden of conventional animal-based production necessitate the development of sustainable and nutritionally balanced alternatives. This review explores the design of a novel hybrid protein powder integrating whey protein—renowned for its superior bioavailability and leucine richness—with corn protein, a sustainable plant-based source possessing the highest leucine concentration among cereals. The innovation lies in employing physical, chemical, and enzymatic crosslinking techniques to form a unified protein matrix with enhanced solubility, stability, and bioactivity, thereby overcoming the intrinsic limitations of individual proteins. The paper critically examines the nutritional rationale underpinning this combination, emphasizing essential and branched-chain amino acids vital for muscle metabolism. It further outlines the extraction and formulation methodologies for both whey and corn proteins, detailing strategies for achieving optimal functional and sensory properties. By merging the anabolic potency of whey with the sustainability and affordability of corn, this hybrid system offers a versatile and eco-efficient ingredient suitable for beverages, bakery products, and fortified foods. Overall, this work highlights hybrid protein technology as a promising pathway toward sustainable protein innovation and future global food security.

Cultured meat is emerging as a new type of food that can provide animal protein in a sustainable way. Many previous studies employed various types of scaffolds to develop cultured meat with similar properties to slaughtered meat. However, important properties such as flavor were not discussed, even though they determine the quality of food. Flavor characteristics vary dramatically depending on the amount and types of amino acids and sugars that produce volatile compounds through the Maillard reaction upon cooking. In this study, a flavor-switchable scaffold is developed to release meaty flavor compounds only upon cooking temperature mimicking the Maillard reaction of slaughtered meat. By introducing a switchable flavor compound (SFC) into a gelatin-based hydrogel, we fabricate a functional scaffold that can enhance the aromatic properties of cultured meat. The temperature-responsive SFC stably remains in the scaffold during the cell culture period and can be released at the cooking temperature. Surprisingly, cultured meat fabricated with this flavor-switchable scaffold exhibits a flavor pattern similar to that of beef. This research suggests a strategy to develop cultured meat with enhanced sensorial characteristics by developing a functional scaffold which can mimic the natural cooking flavors of conventional meat. Flavour properties are often overlooked in meat cultivation strategies and in the development of culture scaffolds. Here, the authors develop a flavour-switchable scaffold for the enhancement of sensorial characteristics of cultured meat.

For the development of innovative foods and nutritional fortification, research into food gel is essential. As two types of rich natural gel material, both legume proteins and polysaccharides have high nutritional value and excellent application potential, attracting wide attention worldwide. Research has focused on combining legume proteins with polysaccharides to form hybrid hydrogels as their combinations show improved texture and water retention compared to single legume protein or single polysaccharide gels, and these properties can be tailored for specific applications. This article reviews hydrogels of common legume proteins and discusses heat induction, pH induction, salt ion induction, and enzyme-induced assembly of legume protein/polysaccharide mixtures. The applications of these hydrogels in fat replacement, satiety enhancement, and delivery of bioactive ingredients are discussed. Challenges for future work are also highlighted.

Demand for a new protein source to replace meat is increasing to solve various issues such as limited resources and food shortages. Diverse protein sources are being developed, but alternative proteins such as plants or insects need to improve people’s perceptions and organoleptic properties. Therefore, cell-based meat research is intensively conducted, and most studies are aimed at scale-up and cost-down via the research of scaffolds and culture media. Here, we proposed a new food by cell powder meat (CPM), which has a high protein content and a meaty flavor. The powder was manufactured 76% more cost-effectively with less serum than the conventional culture medium and without 3D scaffold. Due to its comprehensive characteristics, the potential applicability of CPM in the cell-based meat industry could be expected.

A quarter of the world’s population has no access to safe foods of high quality due to the inability of traditional agriculture to meet the growing needs. Therefore, cultivated meat produced from a large mass of animal cells in a laboratory is becoming a promising alternative to animal products. In this study, we aimed to develop a technology for obtaining a hybrid cultured meat product from rabbit cells, sodium alginate, and sunflower protein, as well as to analyze its morphological and functional characteristics. We used rabbit stem cells isolated from the greater omentum and exposed to lipogenic and myogenic differentiation, as well as rabbit skin fibroblasts. The cells were placed in a hydrogel of sodium alginate and sunflower protein and cultured for 72 h to biofabricate tissue constructs by using 3D bioprinting. Confocal and transmission electron microscopy was applied to analyze the morphological and functional characteristics of the cells in the constructs. Using 3D bioprinting, we obtained tissue constructs of 30×40×3 mm from rabbit cells, sodium alginate, and sunflower protein. According to confocal microscopy, the cells in the tissue constructs remained viable for at least 72 h. Transmission electron microscopy showed that the cells formed tight junctions and were metabolically active for at least 72 h, with fibroblasts secreting procollagen and lipoblasts secreting lipid droplets. The resulting cellular meat was obtained from a combination of fibroblasts, lipocytes, and myogenic cells, as well as two ink components. The cellular meat product was safe and ready for consumption.

In the field of 3D printing, the physicochemical properties of composite inks are pivotal for constructing accurate printing networks. However, the precise fabrication of molded simulants using food 3D printing technology remains a challenging endeavor. The molecular structure and rheological properties of soybean protein isolate (SPI) make it prone to fracture under high shear stress, compromising printing accuracy and stability. This study aimed to address the deficiencies in precision and stability of existing soy protein-based inks for plant-based meats by developing a food composite ink system that incorporated SPI, konjac glucomannan (KGM), and xanthan gum (XG). The new ink system was designed to capitalize on the interactions between proteins and polysaccharides, as well as the synergistic effects of polysaccharides, to achieve high printing precision and the potential for simulant preparation. The addition of KGM and XG to the ink formulation enhanced the shear-thinning behavior, which was more amenable to the printing process, compared to the control group consisting of SPI alone. The SK1X1 composite ink demonstrated a significant improvement in printing precision by 40 % and in printing stability by 59 %, with final values reaching 99.11 % and 98.51 %, respectively. Additionally, hydrogen bonding was identified as a predominant factor in the gel network structure of SPI-KGM-XG composite inks. The self-assembling behavior of KGM-XG with SPI resulted in a robust spatial network structure, which in turn enhanced the thermal stability of the ink. In conclusion, the SPI-KGM-XG blends were determined to be suitable for use as thermo-extruded edible inks, and the synergistic effect of KGM-XG bolstered the gel properties of the hybrid inks, positioning them as ideal candidates for application in the 3D printing of meat simulants.

Abstract Sustainability is defined as balancing environmental, economic and social factors, and various methodologies and tools are available to assess sustainability across sectors and scales. The demand for food has increased due to the increase in the population and the consumption of aquatic food in the world has increased significantly and is growing by an average of 3 % per year, while the population is growing by 1.6 % per year. As the aquaculture industry expands worldwide, it is important to consider the environmental impact of the industry and choose environmentally friendly alternatives to reduce its impact. The aim of this study is to assess the composition of five different fish feeds based on environmental, economic and social parameters using life cycle analysis (LCA), life cycle costing (LCC), social life cycle analysis (S-LCA) as well as technical considerations. The impact of alternatives to the main feed ingredients was analysed separately, while the development of fish feed focused on the protein source used in the feed and the oil used, as halieutic resources are used as raw material in their production, and alternatives are being considered. The best alternatives, considering all four dimensions, were the alternatives where fishmeal protein was partially replaced by Black Solder fly and Yellow Mealworm protein, as the proportion replaced is different for each alternative. By evaluating fish feed along several dimensions, the aim is to improve fish welfare while reducing the environmental impact of feed production.

The trend in the consumption of unconventional, more nutritious foods is leading to the globalization and decentralization of their production, giving rise to the adaptation and innovation of traditional products to make them healthier and more sustainable. This article focuses on quinoa and aims to estimate the environmental impacts of its production adapted to the Spanish conditions and of potential derived snacks enriched with this pseudo cereal by applying conventional and nutritional life cycle assessment (LCA) methodologies. Besides, an exhaustive study of the energy flows by measuring the cumulative energy demand and the calculation of the energy return of investment (EROI) is carried out to assess the most impactful aspect of the processing industries. The application of cradle to gate LCA revealed that polluting emissions of Spanish quinoa are rather similar to those of the Andean grain, with an impact on climate change of 1.03 kg CO2 eq./kg. However, high resource footprints were obtained, for instance a water deprivation potential of 60 m3/kg due to the scarcity in this country. Besides, the consideration of a nutrient profile model as functional unit led to the conclusion that quinoa-based snacks are generally more environmentally sustainable than their conventional counterparts in terms of climate change, resources consumption or water degradation. EROI scores were relatively low for all options, with only between 1.77 and 4.35 % of the energy invested returned, which evidences the unsustainable agricultural practices and low efficiency of processing units. Based on this research, producers can reorient production systems in support of nature, and consumers are able to guide their choices towards improved eating patterns.

The growing global population and unsustainable consumption of natural resources are expected to exacerbate future food security challenges. Cultivated meat, a novel technology produced by culturing animal cells in controlled environments without the need for conventional livestock farming, offers a sustainable alternative that also supports animal welfare. This study presents a literature review focused on the environmental impacts of cultivated meat production, specifically analyzing studies utilizing the Life Cycle Assessment (LCA) methodology. In addition, the review considers health and safety implications, animal welfare concerns, and the economic landscape of the cultivated meat sector. Findings suggest that cultivated meat can be a sustainable solution, particularly when renewable energy sources are employed. However, energy consumption remains the primary contributor to environmental impact, with greenhouse gas emissions largely associated with energy use and culture media. The implications of large-scale production are still uncertain. While cultivated meat may benefit animal welfare, its long-term health effects are not yet fully understood. Despite a decline in investment in 2023, new market entrants and decreasing production costs reflect ongoing industry momentum. Overall, cultivated meat demonstrates strong potential as a sustainable protein source, though further research and technological development are necessary to realize its full promise.

This review examines the history of consumption, life cycle, and culture conditions of seven edible mucilaginous terrestrial cyanobacterial strains—Nostoc flagelliforme, Nostoc commune, Nostoc sphaeroides, Nostoc sphaericum, Nostoc verrucosum, Aphanothece sacrum, and Nostochopsis lobatus—as resilient and sustainable food sources in the face of climate change. Traditionally consumed across various cultures and known for their resilience in extreme environments, these cyanobacteria offer high nutritional value, including proteins, vitamins, and essential fatty acids, making them promising candidates for addressing food security. Their ability to fix nitrogen reduces reliance on synthetic fertilizers, enhancing agricultural applications by improving soil fertility and minimizing dependence on fossil fuel-derived chemicals. Unlike conventional crops, these cyanobacteria require minimal resources and do not compete for arable land, positioning them as ideal candidates for low-impact food production. Despite these advantages, the review highlights the need for scalable and cost-effective cultivation methods to fully realize their potential in supporting a resilient global food supply. Additionally, it underscores the importance of ensuring their safety for consumption, particularly regarding toxin content.

Fusarium species offer a promising solution for cultivating mycoprotein, a sustainable alternative protein source with a variety of food applications. This review comprehensively examines the properties and cultivation methods of mycoprotein production using Fusarium biomass. Fusarium strains' capacity to produce biomass with high protein content, and the ability to utilize diverse substrates, make them ideal candidates for large‐scale cultivation and mycoprotein production. For maximum mycoprotein yields, the parameters that can be optimized for fermentation like temperature, pH, inoculum size, fermentation time, cultivation method, stirring speed, and substrate composition are explored in various studies enlisted here. The different methods of cultivation along with their advantages and disadvantages are also discussed. Furthermore, the nutritional composition and rheological attributes of Fusarium‐derived mycoprotein are reviewed, highlighting its potential as a viable substitute for animal‐based proteins in products like meat and dairy analogues. This review also sheds light on the Fusarium mycoprotein products already in the market and their sustainability values. Overall, mycoprotein derived from Fusarium species is a promising sustainable food ingredient, with opportunities for further improvement in developing more sustainable growth media and enhancing consumer acceptability.

Entomophagy has received widespread attention in both developed and underdeveloped nations. However, the exponential increase in demand for the few available conventional food and feed ingredients that are limitedin supply and production has spurred the interest of food scientists and researchers to scale up their efforts to attain a more sustainable food ecosystem. This review, however, focuses on the current state of knowledge and the prowess of entomophagy to provide global faunas (micro and macro) with more extensive food and feed options. Edible insects potentially satisfy human dietary protein demands compared with other protein sources such as beef, chicken and pork. Unlike other animal-based proteins, edible insects contain higher protein levels, reaching 30% to 85% on a dry matter basis, and can quickly transform low-quality organic waste into edible protein. More so, studies have shown that replacing fishmeal with insect meal is a suitable alternative protein source for farm animals. Insect farming refers to the practice of raising and breeding insects as mini-livestock for the purpose of producing food or feed products that can be consumed directly by humans or animals. Insect farming is environmentally friendly due to its low ecological impact and the minimal need for arable land and water resources compared to other livestock species using about 80% of available agricultural land worldwide. When globally accepted, entomophagy and commercial insect farming have the potential to ease the hunger threat for the predicted world population, expected to be between 9.4 to 10.1 billion people in 2050.