Cutting-Edge Technologies of Meat Analogs: A Review

Current research on cultured meat primarily focuses on reconstructing muscle tissue in vitro; thus, the sensory attributes, such as taste, texture, and nutritional content of cultured meat, are often overlooked. The wide range and availability of meat alternatives in the market are significant for consumers, but they need to meet several criteria, including appearance, flavor (both aroma and taste), and texture. Many consumers place a high value on the taste and texture of meat, making these factors essential for consumer acceptance (Hoek et al., 2011). Actually, the taste and texture there are reported barriers to its acceptance by meat consumers. Trans-differentiation refers to the process of converting mature cells into a different cell type and can be utilized to convert muscle cells into adipocytes. Ma et al. (2024) established a protocol for the controlled trans-differentiation of chicken fibroblasts into myoblasts (Table 1). They confirmed the same results as previous studies that used treatments such as chicken serum medium and fatty acids to induce the conversion to adipocytes capable of fat storage (Lee et al., 2021). Furthermore, using immortalized cells derived from chicken, Ma et al. (2024) exploited the adipogenic/adipogenic switch differentiation of chicken fibroblasts in a 3D culture system utilizing gelatin methacryloyl (GelMA) hydrogel to accumulate fat in muscle tissue without co-culture or mixing of fat. Using fish gelatin/alginate scaffolds, muscle and fat structures were assembled by regulating cell differentiation to produce cultured meat with a softer texture and higher elasticity with a flavor similar to traditional beef (Lee et al., 2024c).

Cultured meat production can be made more effective and efficient by using biomass and fermentation technologies with the aim of improving the texture and flavor of cultured meat (Attaran Dowom et al., 2019). In particular, precision fermentation has improved the organoleptic properties of cultured meat by using genetically engineered organisms to biosynthesize secretory bio-based heme (Jin et al., 2018) and by inserting targeted genes into specific yeast strains and fermenting them to produce fats with a molecular structure similar to that of conventional meat (Singh et al., 2022). Precision fermentation has also been used to produce bitter blockers to eliminate the bitterness associated with soy-based additives, and supernatants of cultures used to ferment cordyceps and several fungal strains (genetically engineered DH5 Escherichia coli) have been added to foods to reduce bitterness (Dunstan et al., 2020). In this way, biomass fermentation using fungi can not only help improve the flavor of cultured meat but also contribute to a sustainable industry by utilizing resources. Texture profile analysis and rheological methods were used to characterize mechanical properties of cultured meat (Paredes et al., 2022). The textural properties (hardness, cohesiveness, chewiness, resilience, springiness, and Young’s modulus) of commercial sausage, turkey breast, chicken breast, and cultured meat were similar, and sausage cooked with cultured meat exhibited higher elastic modulus and shear modulus than other commercial meats, identifying some parameters that should be considered in the initial cultured meat manufacturing process.

Even though cultured meat is not yet commonly available in the market, consumer interest in trying it has been unexpectedly high, and its increasing popularity is promising. Cultured meat presents as feasible path for future development, however, further studies related with the mass production on consumer acceptance would be advantageous to secure success when the cultured meat is widely launched in the future. In the development of cultured meat, stable cell proliferation and optimized culture conditions are key to improving production efficiency (Table 1). As a strategy for stable cell proliferation, immortalization of cells is being investigated. Fibroblasts from several chicken breeds were naturally immortalized to form genetically stable cell lines, which grew at high densities despite the use of serum-free media and were able to produce high yields of cultured meat without genetic modification (Pasitka et al., 2023). Research on optimized culture conditions to improve production efficiency is also ongoing. When Hanwoo myosatellite cells and C2C12 cells were cultured at 37°C and 39°C to compare their proliferation and differentiation efficiency, it was found that culturing at 39°C, which significantly increased the gene expression levels of myosin heavy chain (MyHC), myogenic factor 6 (MYF6), myogenin (MYOG), and myoglobin (MB), could increase the production efficiency of cultured meat (Oh et al., 2023). In addition, the use of pronase is more efficient than collagenase for the efficient isolation of porcine muscle stem cells, specifically the use of pronase and Dispase II, which promotes higher attachment and proliferation rates of muscle stem cells (Li et al., 2022a).

To reduce production costs and address the stability issues of cultured meat, research is being conducted on the development of serum-free media and various additives for the successful development of cultured meat. It has been confirmed that efficient production of high-quality cultured meat is possible by checking the metabolism of muscle cells (Jang et al., 2022). Non-essential amino acids, pyruvate reduction, and transamination showed significant differences in serum and serum-free medium (B27, AIM-V) culture conditions, and the metabolic profile confirmed the predominance of glycolytic and oxidative metabolism in C2C12 myotubes cultured in serum and B27 medium. Dai et al. (2024) selected 19 components and developed an optimized serum-free medium by combining each component through a Plackett Burman design. Through this approach, they obtained a system capable of long-term culture and mass production of C2C12 myoblasts. Beefy-9, a serum-free medium for long-term culture of bovine satellite cells (BSCs), was developed by adding recombinant human albumin to promote BSC growth and maintain myogenicity of the cells and was shown to be suitable for mass production at an economical cost (Stout et al., 2022).

Fibroblast growth factor 1 (FGF1) signaling plays an important role in muscle stem cell proliferation. It is known to contribute to the proliferation of satellite cells and maintain their proliferation rate. Its effectiveness as an additive is excellent, but its high cost is a barrier to cultured meat production. Liu et al. (2024) developed a method to efficiently produce soluble, bioactive recombinant bovine FGF1 (rbFGF1) protein in E. coli. rbFGF1 promoted mitochondrial division and proliferation of MuSCs under serum-free medium conditions. The study showed that rbFGF1 promoted the proliferation of C2C12 cells, increased the mitochondrial membrane potential, and induced cell proliferation through extracellular signal-regulated kinases1/2 (ERK1/2) signaling, making it an effective additive in serum-free medium conditions. FGF2, widely used in cultured meat production, is an expensive growth promoter that increases the cost of serum-free media for related cells, including MuSCs. To lower the cost, one study developed immortalized BSCs (iBSCs) that can grow without external growth factors by overexpressing FGF2 and mutant Ras^G12V (Stout et al., 2024). These immortalized cells were shown to be able to proliferate for multiple generations in FGF2-free medium and retain their originality despite reduced root canal formation, confirming that this is an effective cell engineering technique for reducing cultured meat production.

To investigate the feasibility of utilizing extracted components derived from microalgae as growth promoters in serum-free media conditions, an extract from the microalga Chlorococcum littorale was obtained by sonication. The addition of the extract to mammalian cell lines (C2C12 cell lines, 3T3 cell lines, and CHO cells) in place of fetal bovine serum (FBS) resulted in high proliferation rates (Ghosh et al., 2024). Glucose extracted from C. littorale or Arthrospira platensis and many amino acids extracted from Chlorella vulgaris have been shown to be excellent as media additives for C2C12 mouse myoblast cell cultures (Okamoto et al., 2020). Furthermore, the acid hydrolysis method for extracting the growth-promoting factors from the algae is regarded as simple, economical, and environmentally friendly, with applications in cultured meat production as well as various other fields such as regenerative medicine and gene/cell therapy (Okamoto et al., 2020). Porcine-derived muscle stem cells tend to lose their stemness during cultured meat processing. Therefore, to promote the proliferation and differentiation of the cells, various flavonoids (quercetin, icariin, 3,2′-dihydroxyflavone) were added to the culture media (Guo et al., 2022). While 3,2′-dihydroxyflavone stood out for its role in maintaining stem cell competence, quercetin was superior to the other flavonoids in inducing differentiation and upregulating the expression of MyHC.

Cytokines play an important role in promoting rapid cell proliferation. However, their application in cultured meat production has been challenging due to the high cost of commercial cytokines as well as potential food safety issues (Laulund et al., 2017). Lei et al. (2023) established a system to obtain a recombinant strain CPK2B2 co-expressing four cytokines (including long-chain human insulin growth factor-1, platelet-derived growth factor-BB, basic FGF, and epidermal growth factor) through the Cre-loxP system in Saccharomyces cerevisiae, with a yield of 18.35 mg/L. Subsequent to cell lysis and filter sterilization, the CPK2B2 lysate was used to stimulate porcine muscle stem cell proliferation. Overall, the study afforded a simple and cost-saving approach for cultured meat production and efficient cytokine production.

Scaffolds are often used to maintain the morphology of existing cultures and promote the development of muscle, fat, and connective tissue. Consequently, there is substantial research focused on scaffold formation methods and scaffold materials. To accurately recreate the extracellular matrix (ECM), which regulates the dynamic behavior of cells in tissues, provides structural support, and transmits information between cells, cell scaffold structures must be 3D and contain interconnected networks embedded in viscoelastic materials. Various biomaterials have been designed as in vitro supports that reproduce the ECM to support cell viability, growth, and migration (Tahir and Floreani, 2022). However, the main issue with current scaffolds for cultured meat is striking a balance between the need for food-grade materials and the cell adhesion, proliferation, and differentiation capabilities. Edible proteins have been used as scaffolds to overcome these limitations. Seah et al. (2022) suggested that scaffold technologies for cultured meat should focus on biocompatibility, biodegradability, pore size, strength of the scaffold material, structure, and fabrication techniques. Rabbit skeletal myoblasts (RbSkMCs) cultured on gelatin fibers produced from co-spinning gelatin and microbial transglutaminase (a food-grade crosslinker) formed 3D tissues with visible cytoskeletal (F-actin) networks throughout the tissue (MacQueen et al., 2019). Jiang et al. (2013) demonstrated that electrospun zein nanofibers can be used for cell adhesion and cell growth of fibroblasts. In another example, scaffolds synthesized from gelatin and soymilk expressed myosin in C2C12 cells and upregulate the expression of PPARγ, an adipogenic transcription factor, in 3T3-L1 cells (Li et al., 2022a). It was found that the bioactive isoflavones such as daidzein, genistein, and glycitein from soymilk induced muscle formation, suggesting that abundant integrin binding sites of gelatin help to improve cell adhesion and migration and induce differentiation signaling, which could be useful for cultured meat production (Li et al., 2022a).

Various studies have been published on porous supports that provide space for cell attachment and proliferation. An edible scaffold was developed using various food-grade materials (proanthocyanidins, dialdehyde chitosan, collagen) and various proportions of yeast protein. The developed scaffold had a porous structure and was enriched with tripeptides, which provided space for cell attachment and proliferation, promoting growth and differentiation. In addition, the cultured meat produced through this method showed a level of springiness and chewiness similar to that of traditional meat, which had a positive effect on improving texture (Wang et al., 2024b). Gome et al. (2024) cultured immortalized bovine mesenchymal stem cells (bMSCs) using a macrofluidic single-use bioreactor fabricated with a plant-based scaffold and polyamide polyethylene (PAPE) film as a leak-proof, strongly sealed form of container. The porous structure of rice puffs used as the plant-based scaffold resulted in improved cell attachment and proliferation, and high water absorption was confirmed, indicating that it can be utilized for economical and efficient cultured meat production. Glutenin-chitosan 3D porous supports with pore diameters of 18 to 67 μm and compression moduli of 16.09 to 60.35 kPa effectively promoted cell growth and muscle differentiation and were found to be effective in enhancing the texture and mouthfeel of porcine-derived cultured meat by increasing multiple myofibril fusion with cytoskeleton expansion and tissue maturation (Wu et al., 2024). In addition, customized supports were also developed for fish cultured meat to analyze biological differences. Satellite cells obtained from the large yellow croaker were cultured in two-dimensional (2D) and 3D cell culture systems. The results showed that cell adhesion receptors and myogenic markers [paired box 7 (Pax7), myogenic differentiation 1 (MyoD1), desmin] were more highly expressed in 3D cultures using hydrogel/microcarrier, which was favorable for the production of fish cultured meat (Yin et al., 2024).

Alginate, which is often used in scaffolds, is derived from brown algae and is known to be sustainable, readily available, and an ideal material for making hydrogel supports (Kang et al., 2021). One study synthesized methacrylated alginate (AlgMA) through a dual crosslinking system to form covalent bonds and arginyl-glycyl-aspartic acid (RGD) conjugates (AlgMA-RGD) to develop hydrogels with tune bale mechanical properties (Tahir and Floreani, 2022). The hydrogel scaffolds showed excellent effects on the viability and attachment of MuSCs. Jeong et al. (2024) developed a technique for culturing and aligning muscle cells using zein-alginate fibers made by coating zein protein on alginate fibers. The fibers were produced using a wet spinning technique that takes advantage of the fact that when zein solution encounters hydrophilic alginate hydrogels, its solubility decreases, and it coagulates on the alginate surface. The fibers had excellent cell affinity, biodegradability, and a high strain rate of more than 75%, which promoted the maturation of muscle cells and the formation of aligned myotubes. This economical and simple production of fiber scaffolds using plant-based materials can eliminate toxic chemicals as a limiting factor and is conducive to cell stacking and structuring, suggesting the potential for mass production of cultured meat.

Depending on the material and combination of inks, 3D printing can affect the texture and various rheological properties, which can significantly impact quality. 3D printing could provide solutions for the critical issues of cultured meat, utilizing by-products and solving sustainable industrial and food contamination problems (Dong et al., 2023). A 3D printable scaffold based on gelatin/alginate/ε-poly-L-lysine hydrogel was developed to provide a platform for cultured meat production by confirming the cell attachment, increased proliferation rate, and maintenance of the differentiation capacity of C2C12 mouse skeletal myoblasts and porcine muscle stem cells (Wang et al., 2024a). In addition, a polydimethylsiloxane mold fabricated by 3D printing technology was used to form a 3D skeletal muscle tissue network using porcine muscle stem cells (Zhu et al., 2022). The results showed improved texture and increased amino acid content, suggesting an effective working process for cultured meat production. Another study generated a meat-like cell sheet structure without 3D constructs, bioprinting technology, or ECM components for cell attachment. The meat-like tissues with meat-like texture could be produced by combining myoblasts (C2C12) and adipose progenitor cells (3T3-L1) in an optimal ratio (Shahin-Shamsabadi and Selvaganapathy, 2022).

Research is also being conducted on the aspects of cultured meat safety and health protection. Atlantic mackerel (Scomber scombrus) skeletal muscle cells were used to determine the effects of microplastic exposure on the production efficiency of cultured seafood (Sun et al., 2024). The results showed that microplastic concentration significantly affected cell adhesion and proliferation but not differentiation. There have also been studies that have used a combination of different techniques to produce cultured meat. Porcine pre-gastrulation epiblast stem cells were used to develop a cell line capable of long-term culture and genetic stability, and a differentiation system was established with serum-free medium to successfully differentiate muscle cells. Edible scaffolds with porous and homogeneous structures were utilized to produce cultured meat through ionic crosslinking of various formulations of konjac glucomannan, sodium alginate, and calcium ions (Zhu et al., 2023).

The market for edible insects is forecasted to grow to $17.9 billion by 2033, with a CAGR of 28.6% from 2024 to 2033. In terms of volume, it is anticipated to reach 4.7 million tons by 2033, growing at a CAGR of 36.3% over the same period (Meticulous Research, 2023). Insect protein is one of the emergent protein sources expected to enter the sustainable food market and is a highly efficient protein source. Insect farming is an environmentally friendly way of raising animals, with less water and land requirements than farmed animals and less pollution compared with the pollution issues associated with livestock farming. Insect-derived ingredients are mainly composed of proteins derived from insect muscles, fat bodies, and the cuticle layer that covers the epidermis (Lamsal et al., 2019). Insect-based protein ingredients are often made by grinding whole insects and processing them into powder for various food or feed applications, with insect protein content varying from 13% to 77% by building (Lamsal et al., 2019). Edible grasshoppers in Mexico exhibited a protein content of 44% to 77% and a fat content of 4% to 34%, which fulfills essential amino acid requirements (Blásquez et al., 2012; Paul et al., 2016). Defatted, alkaline, and ultrasound-assisted extraction methods were found to increase the amount of protein extracted from edible mealworm (Schistocerca gregaria) and honeybee (Apis mellifera) by 57.5% and 55.2%, respectively (Mishyna et al., 2019a; Table 2). Furthermore, the extracted proteins exhibited high foaming and emulsification stability, and the proteins obtained from honeybees, in particular, exhibited high thermocoagulability, indicating that insect-based proteins have the potential as food feeds or dietary supplements (Mishyna et al., 2019a). Similarly, Queiroz et al. (2023) found that techniques such as autoclaving, sonication, pulsed electric fields, and ohmic heating improved various functional properties of insect proteins, including solubility, emulsification, and foam-forming ability. Another study investigated the temperature- and pH-dependent aggregation and gelation of edible bee larval proteins (Mishyna et al., 2019b). The highest aggregation rates (73.7% and 68.4%, respectively) were found at pH 5 and 7 at 85°C, and the protein properties were changed by heating, with minimum gelling concentrations of 5% at pH 7% and 11% at pH 3. Therefore, the potential of using edible insects as gelling agents was confirmed. Lee et al. (2024b) confirmed that Tenebrio molitor larvae ethanol treatment could change the structure of proteins to improve their technical and functional properties. They found that the protein molecular weight decreased, the α-helix structure decreased, and the β-sheet structure increased with increasing ethanol treatment concentration. In addition, ethanol treatment increased protein surface hydrophobicity and foaming ability, and the best antioxidant activity was found at 20% ethanol treatment (Lee et al., 2024b).

The development and evaluation of edible insect-based food products are being actively explored. The addition of insect protein (Alphitobius diaperinus) was shown to decrease the pH, monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) while increasing total lipids and saturated fatty acids (SFA) in burgers, with the highest sensory acceptability score at 5% insect protein (Krawczyk et al., 2024). A study on the preparation of baby biscuits using wood grasshopper flour (Melanoplus cinereus) found that the addition of 5% insect powder provided the most appropriate energy and nutrient content and was the most preferred in terms of taste, aroma, and texture (Dewi et al., 2020). In addition, the developed biscuits also provided high-quality protein and were found to meet Indonesian supplementary food regulations. The effect of insect addition, extrusion temperature, and moisture content on the texture properties of a meat substitute prepared from cricket flour and soy protein isolate was determined using a high-moisture extrusion cooking technique (Kiiru et al., 2020). The tensile strength increased with increasing extrusion temperature but decreased with increasing cricket flour addition. In particular, meat alternatives with the best properties and texture were obtained at an extrusion temperature of 160°C and 30% low-fat cricket flour addition, and moisture content was found to play an important role in the structural integrity and texture formation of meat alternatives (Kiiru et al., 2020). Totally, Queiroz et al. (2023) reported ultrasound processing, high hydrostatic pressure, pulsed electric fields collectively provide innovative solution for improving the textural and flavor profiles of insect-based protein products, making them more appealing as meat alternative ingredients. Insect protein is a highly sought-after resource due to its high protein and oil content. However, it has very low consumer acceptance, so many researchers have been adding insects in powder form to existing foods or processing them as a way to address this issue. Furthermore, to maximize the potential of insects as an alternative food resource, it is believed that research should focus on demonstrating the functional properties (water and fat absorption capacity, emulsification, foaming, gelation, rheological properties) and various food applications of insect protein.

Based on market analysis, the US mycoprotein industry is expected to achieve a value of $523.4 million by the end of 2030 (Khan et al., 2024). In comparison, China’s market potential is anticipated to reach approximately $238.6 million by 2030 (Derbyshire and Delange, 2021). Additionally, this figure is expected to exceed $1.1 billion by 2030. Mycoprotein contains high-quality protein, 25% fiber, high amounts of MUFA and PUFA, low SFA, a wide range of essential amino acids, and produces less greenhouse gas emissions than plant and animal proteins (Ahmad et al., 2022; Khan et al., 2024). It also contains similar levels of protein utilization to milk-derived protein and has been shown to be beneficial for muscle protein production in children (Monteyne et al., 2020). Mycoprotein is primarily produced through a fermentation process, although it can also be produced from various agricultural wastes (Zhang et al., 2023). It is currently manufactured by Marlow Foods under the brand name Quorn by culturing Fusarium venenatum in a specific environment and mixing it with egg whites, colors, and flavoring compounds to create a meat-like texture, which is sold in the United States, United Kingdom, France, Europe, and Singapore (Finnigan et al., 2019; Khan et al., 2024). Fermentation methods for the mass production of mycoprotein are classified solid-state fermentation (SSF) and submerged fermentation (SmF; Ahmad et al., 2022; Majumder et al., 2024). SSF has been utilized for centuries in the production of traditional fermented foods like soy sauce, tempeh, miso, koji, red fermented rice, and tapai (Lizardi-Jiménez and Hernández-Martínez, 2017). Mycoprotein produced via SSF, which operates under low moisture conditions and uses solid substrates, is nutritionally superior, with a higher protein digestibility-corrected amino acid score than chicken or beef, effectively addressing the limitations of plant-based proteins (Cerda et al., 2019; Kim and Kim, 2012). SmF involves the growth of microorganisms in a liquid medium with high water content (over 95%), utilizing carbon, nitrogen, and micronutrients for cultivation (Majumder et al., 2024). This method, which supports both anaerobic and partially anaerobic processes, has become a preferred technique in the production of bioactive mushroom compounds due to its ease of management and rapid control of growth conditions (Majumder et al., 2024). The SmF process for mycoprotein production includes microbial growth in the liquid medium, followed by incubation, centrifugation, washing, and filtration to isolate the protein-rich biomass (Reihani and Khosravi-Darani, 2019). While SmF is widely used in large-scale industrial enzyme production due to its efficient handling and monitoring capabilities, it is also associated with significant challenges, including high costs, lower productivity, and the complexity of the required medium (Ahmad et al., 2022; Manan and Webb, 2017).

Pavis et al. (2024) found that a dietary invention with mycoprotein-containing foods significantly reduced total cholesterol, low-density lipoprotein cholesterol, and non-high-density lipoprotein cholesterol concentrations in adults with overweight, improving cardiometabolic health by reducing hypercholesterolemia. Neurospora crassa mycoprotein, traditionally used as a fermented food mainly in Indonesia and China, has been safely used as animal feed (Liu et al., 2016). This nutritious material is rich in complete protein (45 g/100 g), dietary fiber (35 g/100 g), potassium, and iron with no toxicity or allergenicity, confirming its potential as an alternative material for meat production (Bartholomai et al., 2022). Nuggets prepared using mycoprotein were found to have 57.9%, 24.1%, 13.2%, and 2.1% moisture, protein, fat, and ash content, respectively, and exhibited similar characteristics to chicken nuggets in terms of texture, color, and physicochemical properties, and with 33% lower cooking loss (Hashempour-Baltork et al., 2023).

The main sources of plant protein are grains (wheat, corn, oat, rice, rye), legumes (pea, red bean, chickpea, lentil, faba bean), seed oils (sunflower, rapeseed, flaxseed, hemp seed, cotton seed, sesame seed, pumpkin seed), nuts (almond, pistachio, cashew, walnut, peanut), and others (quinoa, buckwheat, chia seed, amaranth, potato; Munialo, 2024). Alternative proteins, such as plant-derived proteins, have the potential to contribute to reducing the environmental impact of animal agriculture. While plants produce much lower greenhouse gas emissions and require fewer resources than animals, structural changes to water-soluble proteins during extraction or drying can affect the protein’s function and bioactivity (Munialo, 2024). Plant-based proteins are also often associated with a lack or imbalance of essential amino acids, reduced flavor and texture compared to animal proteins, and potential allergenicity, which can be a major barrier to consumers choosing plant-based protein products. Therefore, there is a need to address taste, nutritional value, and allergenicity to create meat alternatives using plant-based proteins.

In order to utilize plant proteins as a substitute for animal proteins, it is important to select a suitable extraction method based on the protein matrix. Protein extraction methods include dry, wet, enzymatic, sub- and supercritical water extraction, and reverse micelles extraction (Thakur et al., 2024). Dry methods utilize air and are energy efficient; however, they are known to be unsuitable for certain feedstocks due to their lipid content, which can lead to fractionation or cluster formation of impurities (Banjac et al., 2017). Wet methods can utilize different types of extraction solvents, and functional properties such as gelation, foaming, and emulsification can be applied to the extraction method. Methods that utilize enzymes to degrade proteins and cell walls/membranes require long processing times, high costs, and high energy consumption but can minimize environmental impact and yield high-quality samples (Gouseti et al., 2023; Sari et al., 2013). Subcritical-supercritical extraction does not require organic solvents and is a sustainable extraction method, with protein content increasing with increasing temperature (Knez et al., 2018). Finally, reverse extraction can be significantly affected by pH changes, electrostatic interactions, concentration and nature of the target protein, ionic strength, and reverse micelles composition (Zhao et al., 2018).

Physical, chemical, and biological methods can be used to improve the functionality of vegetable proteins to increase their utilization or incorporation into various products, especially by enhancing their taste, nutritional, and functional properties (Nasrabadi et al., 2021; Table 2). Nowacka et al. (2023) identified various chemical (glycosylation, deamidation, phosphorylation, acylation), physical (pulsed electric fields, ultrasound, high hydrostatic pressure, dynamic high-pressure treatment, cold plasma), and biological (fermentation, enzymatic modification) methods that are used to enhance the functional properties of proteins. They suggested that meat- and fish-like products can be produced by appropriately combining different production techniques and materials (Nowacka et al., 2023).

A growing body of studies is exploring the utilization of plant-derived proteins to develop meat substitutes by identifying their functional properties and analyzing their structure and texture. Plant-based protein products as substitutes for animal protein include plant-based meat, plant-based eggs, plant-based dairy products, and plant-based protein emulsion foods, which can be manufactured by techniques such as high-moisture extrusion, 3D printing, and electrospinning (Xiao et al., 2023). In a previous study, pea and rice were selected as low-allergenic materials analyzed for their functionalities in comparison to wheat and soybeans (Zhao et al., 2020). It was found that although the functionality of pea protein is similar to that of soybean protein and superior to rice protein, the excellent water absorption and emulsification properties of rice protein make it suitable as an alternative protein material. To evaluate the potential of banana floret (Musa paradisiaca) and jackfruit (Artocarpus heterophyllus Lam.) as meat substitutes, they were incorporated into a vegan sausage (Keerthana Priya et al., 2022). The developed vegan sausage was rich in fiber and protein. It showed good hardness, adhesion, chewability, and elasticity, improving the texture properties and overall palatability, which was similar to commercial chicken sausage. Oat fiber concentrate and pea protein isolate were processed using a high-moisture extrusion method to prepare fibrous meat substitutes with 33% dietary fiber content (Ramos Diaz et al., 2022). An evaluation of the mechanical and physicochemical properties of the meat substitutes showed that the structure became softer with increasing dietary fiber, the high-cooling temperature strengthened the fiber structure, and the extractability and viscosity of β-glucan were well maintained using a human digestion model. In another example, a biopolymer composite with a meat-like texture was fabricated by complex aggregation of gellan gum, a polysaccharide, and potato protein, and the electrical characterization, microstructure analysis, dynamic shear flow, and texture of the composite were analyzed (Hu et al., 2024). At pH 4, excessive air bubbles formed after heating, resulting in a spongy gel with relatively low hardness. In contrast, at pH 6, a fibrous structure with the highest hardness and elasticity was formed after heating. Additionally, in the high-moisture extrusion process, disulfide bonds significantly contribute to the fibrous texture of meat analogs, largely due to the important role of wheat gluten (Chiang et al., 2019). By employing a Shear Cell or a lab-scale Couette Cell, highly anisotropic fibrous samples can be produced under processing conditions. Couette cell is preferred for its scalability and potential for future continuous operation, which facilitates the production with uniform thickness, while the Shear Cell is more suitable for laboratory-scale applications (Krintiras et al., 2016). Therefore, it was suggested that composite aggregation and thermal coagulation will help in the formation of plant-based products with meat-like texture and structure.

Plant-based protein products are important not only for vegetarians but also for those who want to eat a low-fat, high-fiber diet to reduce the risk of disease and decrease their intake of calories. A study by Meixner et al. (2024) examined where plant-based protein products provide the highest value utility in each food market and how much Generation Z consumers are willing to pay for them. The most important attributes of plant-based protein products were origin, price, and vegan, in that order, with domestic and European products being more positively valued than third-country imports. To increase consumer awareness of plant-based protein products, the first step is to create a favorable product. The selection of optimal plant-derived ingredients and processing technologies that can improve the functional properties of high-quality proteins is crucial. It will contribute significantly to the growth of environmentally friendly, nutritionally rich, sustainable meat substitutes.