P U B L I C A T I O N S CODON Italian Journal of Food Science, 2022; 34 (2): 9–20 ISSN 1120-1770 online, DOI 10.15586/ijfs.v34i2.2141 9 P U B L I C A T I O N S CODON Effect of high-moisture extrusion on soy meat analog: study on its morphological and physiochemical properties Monirul Islam1,2, Yatao Huang1, Md. Serajul Islam3, Ningyu Lei1, Lei Shan1, Bei Fan1, Litao Tong1, Fengzhong Wang1* 1Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-Products Processing, Ministry of Agriculture and Rural Affairs, Beijing, People’s Republic of China; 2Rural Development Academy (RDA), Bogura, Bangladesh; 3State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China *Corresponding Author: Fengzhong Wang, Institute of Food Science and Technology, Chinese Academy of Agricul- tural Sciences, Beijing, 100193, People’s Republic of China. Email: wangfengzhong@sina.com Received: 10 October 2021; Accepted: 7 December 2021; Published: 2 April 2022 © 2022 Codon Publications OPEN ACCESS PAPER Abstract There has been a growing interest in meat analog, microstructure characteristics, and anti-nutritional content obtained from soybean. High-moisture extrusion parameters are the input extruder of moisture content (>40%) that get the advantages of lower energy input. Thermo-mechanical treatment has a considerable influence on structural properties of soy-based meat analog. Texturized soy proteins can substitute meat products while pro- viding a high-protein food ingredient which can be consumed directly as meat analogs. Therefore, this review aims to the effect on soybean of micro-structural and physicochemical properties of meat analogs by high-moisture extrusion. Thus, further studies are required concerning a large-scale meat products with purify protein structure. Keywords: soy meat analog; high-moisture extrusion; allergenic protein; anti-nutritional factors; textural properties Introduction Soybean (Glycine max), which has its origin in East Asia (especially in China and Japan), belongs to the legume family (Kader et al., 2017). It is an excellent alternative source of proteins, complex carbohydrates, polyun- saturated fatty acids, soluble fibers, and isoflavones. Nowadays, it’s grown worldwide as an edible bean (Peluso et al., 2014). Wang et al. (2020) reported that 60 varieties were used for the product development by 2018; 398 mil- lion tons of soybeans were produced worldwide, with 61% for oilseed production. In addition, Salgado and Donado- Pestana (2011) found that 90% of the total soybean pro- duction in the world is from the United States of America (USA), Brazil, South America, and Northwestern Europe. Moreover, Jooyandeh (2011) reported soybean oil com- position that is 15% saturated, 61% polyunsaturated, and 24% monounsaturated fats. Furthermore, Fiala (2008) described that by 2030, the increase in the population, industrialization, and urbanization will also increase the plant meat demand by about 72%. Golbitz and Jordan (2006) found that soybeans typically contain 35–40% proteins with well-balanced amino acid composition, 30% carbohydrate, 15–20% fat, 10–30% moisture content as well as fiber, calcium (Ca), iron (Fe), zinc (Zn), and vitamin B complex. The nutritional composition of soybean/legume is presented in Table 1. In addition, soybean contains some minor compounds, such as lecithin, isoflavones, bio-peptides, and others, that are effective against chronic cancer, cardiovascular diseases, and type II diabetes (Dixit et al., 2011; Singh, 2010; Singh et al., 2008). Soybean protein is suitable for people, who lack plant protein, for nutritional value and mailto:wangfengzhong@sina.com 10 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. Table 1. Nutritional composition of plant based protein, Quorn protein and microbial proteins legume. Protein source Protein content Mineral content References Legume protein Legume seed (20–30%) – (Riascos et al., 2010) – Soy protein (concentrate): Potassium (2.4%), Phosphorus (0.9%), Calcium (0.4%), Magnesium (0.3), Iron (0.02%), Zinc (0.005%) (Singh et al., 2008) Oilseed protein Safflower seed (13–17%) – (Asgar et al., 2010) Cottonseed (23%) – Rapeseed (25%) – Gourd seed Calcium (0.055%), Magnesium (0.720%), Potassium (0.965%), Iron (0.021%), Zinc (0.016%) (Olaofe et al., 1994) Pumpkin seed Calcium (0.072%), Magnesium (0.778%), Potassium (1.127%), Iron (0.027%), Zinc (0.017%) (Olaofe et al., 1994) Cereal protein Maize (8.8–11.9%) – (Orcutt et al., 2006; Riaz, 2004) Wheat (8–17.5%) – Oats (8.7–16%) – Rice (7–10%) – Barley (7–14.6%) – Rye (7–14%) – Quorn protein Mycoprotein 11% Sodium (mg) 5.0 Cholesterol (g) 0 Iron (mg) 0.5 Zinc (mg) 9.0 Selenium (µg) 20 (Finnigan et al., 2019) Bacterial protein Bacillus cereus (Ram horn) 68% Bacillus licheniformis (Potato starch processing waste) 38% Corynobacterium ammoniagenes (Glucose + fructose) 61% Escherichia coli (Ram horn) 66% Corynobacterium ammoniagenes (Glucose + fructose) 61% Cupriavidus necator (Synthetic growth medium) 40–46% – (Ritala et al., 2017) good health effects on calcium metabolism and lowering cholesterol (Shih et al., 2016). Moreover, soy products, such as soy flour, soy milk/powder, soy protein isolate (SPI), soy protein hydrolysates (Guo et al., 2018; He et al., 2019; Simmons et al., 2012), could be used as food addi- tives and nutraceutical ingredients. Meat alternatives could be classified as plant-based (soy, gluten, pea, etc.), cell-based (vitro or cultured meat), and fermentation-based (mycoproteins) (Sha & Xiong, 2020). In recent years, plant-based meat has been developed for meeting consumer demands, exponentially grown market, and the sustainability of future food supply (Sha & Xiong, 2020). The plant-based meat market’s expan- sion is predicted to increase from $4.6 billion in 2018 to $85 billion in 2030, and as a milestone by 2026, it would achieve $30.9 billion (Sha & Xiong, 2020). Among the several mechanistic techniques for the texturization of plant-based meat, extrusion is the most often applied one (Dekkers et al., 2018). Plant proteins represent a safe, sustainable, and practical non-pharmacological approach for lowering cholesterol. Recently, the most famous attributes of plant-based pro- tein, that is soybean protein, is their health benefits linked to the prevention and treatment of many chronic dis- eases. Regular consumption of soy products reduces one’s risk for chronic diseases such as cancer, heart disease, and stroke (Jooyandeh, 2011). Moreover, plant-based meat analog consumption is also beneficial in protecting against heart disease, lowering blood cholesterol; reduc- ing the risk of cancer, and increasing bone mass (Joshi & Kumar, 2015). Furthermore, recent studies have well established that the plant-based food and beverage help Italian Journal of Food Science, 2022; 34 (2) 11 Effect of high-moisture extrusion Plant-based meat analogs Meat analog is also called meat substitute, faux meat, mock meat, or imitation meat. Analog can be defined as the compound of the same structure, such as texture, but slightly different in composition. There are two stages in the production of conventional meat analogs: prepara- tion of emulsion and formation of its chunk. Typically, the emulsion is primed by mixing, chopping, and emul- sifying proteins, fats, salts, and other inclusions to form a protein matrix. Meat analogs can be produced at low moisture (<35%) using a single screw extruder or at high moisture (>50%) using a twin-screw extruder (Lin et al., 2000). Moreover, Riaz (2004) described that meat analog could be formed into strips, sheets, patties, disks, and other shapes. It can absorb water at least three times its weight when cooked in boiling water for at least 15 min. The fibrous, anisotropic structure of meat analog prod- ucts contributes to the meat-like feel and sensory view (Elzerman et al., 2013). Table 2 presents summaries of the comparison between the plant-based and animal-based alternatives to meat. Meat analogs look like textured meat and are healthy (cholesterol-free) and low-cost. In addi- tion, mycoprotein meat analog, originated from fungus, is used as a healthy food alternative for its high-protein in the improvement or management of immune system, have potential antimicrobial effects, helps in reducing risk of cardiovascular and gastro- intestinal diseases with improved physiological functions, decreases risk of low bone mass as well as very high levels of antioxidant activ- ity (Paul et al., 2019). In addition, high-moisture extrusion (HME), over 40% of moisture contents during processing, has the enormous advantage of lesser energy input, lower waste discharge, higher efficiency, and more excellent value of texturized products. Therefore, it is lately considered as the better choice for developing plant protein-based meat sub- stitutes (Zhang et al., 2019). For texture optimization, exogenous polysaccharides are one of the functional pri- mary additives used in food industries. At the time when proteins are denatured during the extrusion process, the dormant reactive sites of the interior proteins would become available, and the structure proteins would become flexible, which permits the protein-polysaccha- ride interactions. Polysaccharides could be used as cross- linker to alter the conformation of proteins, interact with proteins by cross-linking to protein side chains through Maillard reaction, and form a complex structure of pro- tein (Caillard et al., 2010). Table 2. A comparison among cultured meat, plant-based meat, and animal-based meat. Dimension Cultured meat Plant-based meat Animal-based meat References Background information Origins, history, and technical operation Idea articulated around 1930. Since 2000s, research into animal-cell culturing to produce meat. Available in the market for several decades, made from concentrated protein- soy, wheat, pea by extrusion or coagulation. Traditional in China, Japan, research on cell cultures for various production purposes since early 2000s. (Bryant & Barnett, 2018; Sharma et al., 2015; Smetana et al., 2015) Nutritional value Identical of regular meat. Amino-acid profile nutritionally to meat. Rich in protein but extraction needed due to indigestible walls. Consumption and production rates and patterns Not available in the market (optimists predict market introduction by 2019). Established. Modest, relatively stable market share, increase since by 2010. Products from extracted protein. Position as meat alternative Plant meats are more sustainable and animal friendly than animal-based meat. Functional equivalent to meat. Potentially protein- rich source for human consumption. Products rising. Current practice and situation Technological Proof of principle in 2013. Ongoing research and development. Use of purified ingredients, increased efforts to improve meat similarity. Growing and processing under research, have many challenges. (Berghout et al., 2015; Dekkers et al., 2018; Osen et al., 2014; van der Weele et al., 2019) Lifestyle and Consumption Proper meat produced without animal suffering. Established as vegan alternatives replacing meat. Novel: green product color and market profile. Supporters and opposition Attracts venture capitalists and entrepreneurs in Silicon Valley, animal welfare organizations, and other innovators. Emerging interest from meat industry. Mix of alternative retail sector, experiments with hybrids of meat also startups and meat companies. Pioneer companies and scientists, no consolidated coalition. (Aiking, 2011; Aiking et al., 2006; Alexandratos & Bruinsma, 2012) 12 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. and low-fat content, and quality texture. The primary function of meat analogs is to replace the animal meat in the human diet. The consumers of soybean-based meat analogs include not only vegetarians but also the non-vegetarians who want to reduce their meat con- sumption for health reasons. For example, soybean meat analogs contain protein (50 to 95%, dry matter), and soy protein is primarily used as the protein ingredient (Chen et al., 2010). Samard et al. (2019) demonstrated the high-moisture meat analogs are coupled with a higher integrity index and they have the stability of springiness as well as cutting strength than low-moisture meat sub- stitute which is produced using the similar formula and screw-speed. The mechanical approaches summarized to make plant-based meat analogs are displayed in Table 3. High-moisture extrusion of soy meat The HME approach can be used to impart a specific tex- ture to proteins such as soy-proteins, whey-proteins, pea-protein, or wheat-gluten (Pietsch et al., 2017; Wolz et al., 2016). HME is a crucial technology for trans- forming plant-based protein into palatable meat-like products. During the HME process, the input extruder of moisture content is more than 40%, which results in the advantages of lower energy input and higher qual- ity of the texturized products (Zhang et al., 2019). Chen et al. (2010) described that when moisture content of soy meat is increased from 28 to 60% (wet base), the indica- tor residence time and specific mechanical energy (SME) are significantly decreased. In addition, when cooking temperature is increased from 140 to 150°C, the in-line viscosity at die and SME reduced considerably, partic- ularly at lower moisture contents (LMC). Most studies found the effects of processing parameters were done for low to moderate moisture processing. Still, these parameters can significantly affect protein structure during high-moisture processing (Palanisamy et al., 2019). Due to its desirable texture and nutritional value, soy protein can be used to make soy protein products by the extrusion process (Wu et  al., 2019). For the tra- ditional extrusion methods, extrusion at higher mois- ture conditions would reduce viscous dissipation at the lower extrusion temperature. Still, these changes would be expected during high-moisture extrusion conditions (MacDonald et al., 2009). Zahari et al. (2020) reported the chosen parameters including concentration (0, 20, 40, 60%) of hemp protein concentrate (HPC), target mois- ture content (65, 70, 75%), temperature (40–120°C), and screw speed (300–800 rpm), whereas, SPI was extruded with 500 rpm at 70 and 75% of target moisture content. To summarize some literature, extrusion parameters and selected formulations of high-moisture meat ana- logs are shown in Table 4. MacDonald et al. (2009) also reported that high-moisture extrusion was useful to gen- erate high-quality protein foods. Mechanical treatment is more effective during the high-moisture extrusion pro- cess for other plant proteins when compared with wheat gluten. Therefore, the influence of HME processing on the change of protein-protein interactions is used to form the anisotropic structures of SPIs (Chen et al., 2011; Fang et al., 2014). Die-cutting Die-cutting is a critical technology used to bring out the indentation of die-cutting of the surface to retouch the processing equipment, where this technique is a complex core unit. The die-cutting indentation position must be precise to cause simultaneously, and the complete plat- form stress must assure that the place of two active faces in contacts top and bottom to be mutually parallel. Shen et al. (2012) reported the speed of 7000 RH-1, travelling schedule 61 mm, cap board located 80°, the cam midpoint distance was 279 mm in a stand die-cutting machine. For instance, “the follower maximum pivot angle is 20°, main cam swing follower 240-mm long, roller radius 15  mm, vice-cam swing follower 240-mm long, roller radius 15 mm, the base circle initial radius is 60 mm. Moreover, the cam angle for outer dwell is 110°, the cam angle for inner dwell is 10°, work travel angle of follower is 150°, return travel angle of follower is 90°, allowed pressure angle of actuating travel is 35° allowed pressure angle of return travel is 60°, and allowable curvature radius of the real contour line of cam is 3 mm in a platform die-cutting machine (Shen et al., 2012). Table 3. Overview of mechanistic techniques to make plant-based meat analogs. Technique Starting material Equipment type Product Process Key technology References Bottom-up strategy – – Structure/ structural element – Length scale anisotropy (Post, 2012) Wet spinning Protein isolate, coagulation bath Wet spinning setup: barrel, spinning nozzle, water bath, winding device Fibers – Micrometer (Post, 2012) Italian Journal of Food Science, 2022; 34 (2) 13 Effect of high-moisture extrusion the oxidation of polyunsaturated fatty acids, and also may also be responsible for turning into volatile off-flavors. Characteristics of soy meat Microstructure Scanning electron microscopy (SEM), light microscopy (LM), Fourier transform infrared spectroscopy (FT-IR), as well as differential scanning calorimeter (DSC) are usually used to obtain more detailed information on the protein network microstructure of the meat analog. The micro-extraction technique is an environment-friendly procedure due to the reduction of polluting solvents and sample volume. For instance, the micro-extraction approach increases the extraction yield and diminishes the sample equilibrium time (Barzegar et al., 2019). Micro-extraction includes several performances such as single drop micro-extraction (SDME), dispersive- liquid–liquid micro-extraction (DLLME) and hollow- fiber micro-extraction (HFLPME). Preece et al. (2017) reported “the cotyledon-cells protein bodies found in size range from 2.4 to 13.5 μm when used SEM approach with- out sample-hydration.” Lakemond et al. (2000) described two significant storage proteins that compose 60–80% of the total soybean protein: the β-conglycinin and glycinin. Soybean protein is a complex mixture containing various Table 4. Extrusion-parameters involved for screening and selected formulations of high-moisture meat analogs (HMMA). Formulation Speed of screw Targeted moisture content (%) Temperature (oC) (Zone 1–2–3–4) Visual appearance color References 0% HPC and 100% soy SPI 500 70 40–60–80–100 Light color, compact Zahari et al. (2020)500 75 Light color, compact 20% HPC and 80% soy SPI 800 65 40–60–80–100 Compact Zahari et al. (2020)800 70 Compact 800 75 Less compact 600 75 Less compact 600 80 – 400 75 Less compact 300 75 Less compact 40% HPC and 60% soy SPI 800 65 40–60–80–100 Compact, dark spot Zahari et al. (2020)800 70 Compact 800 75 Pale, less compact 60% HPC and 40% soy SPI 800 60 40–60–80–100 – Zahari et al. (2020)800 62.5 – 800 65 Less compact 800 70 Less compact 800 75 Came out foamy 800 65 60–80–100–120 Less compact HPC: hemp protein concentrate; SPI: soy protein isolate. Die pressure The viscosity of the molten blend may be attributed to the decrease in pressure with the increase in temperature. The growth in feed moisture and die temperature then affects the mass thickness over the extruder and decreases the die-pressure value. Zhang et al. (2020a) found that the coefficient variation of the die pressure and die temperature was 26.71 and 1.74%. In addi- tion, the measured die pressure in extrusion cooking of apple meat blend ranged from 9 to 16 MPa. The nega- tive coefficient of the first-order term of temperature, screw speed, and moisture content indicated that die pressure increased with temperature decrease (Singha & Muthukumarappan, 2017). High-pressure homogenization High-pressure homogenization (HPH) is an applicable unit operation based on cavitation to improve processed soybean materials’ extraction yields. Debruyne (2006) demonstrated that homogenization could cause a neg- ative effect on separation efficiency. In addition, inves- tigations may require evaluating the scalability of this promising result obtained by using a lab-scale homog- enizer. Denaturation of the lipoxygenase occurred of enzyme high-temperature treatment may also catalyze 14 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. Tehrani (2014) found that adding soy-flour, flour of split- pea, and wheat-starch could improve low-fat hamburger’s texture properties by reducing shrinkage. For example, Smith et al. (1976) proved that the presence of textured soy protein was associated with the substantial reduction of shrinkage in the blended ground. The transversal cut- ting strength of texturized vegetable protein (TVP) and meat samples was slightly higher than their longitudinal cutting strength. It is stated that the cutting strength in parallel and vertical directions of extrudates could indi- cate the texturization degree or fibrous-structure for- mation (Fang et al., 2014; Gu & Ryu, 2017). When SME is decreased, there is an increase in instrumental chew- iness and hardness. As a result, instrumental chewiness and hardness of meat analogs increased with decreasing SPC-WG ratio (Fiorentini et al., 2020). Additionally, Fang et al. (2014) reported that the instrumental hardness and chewiness of texturized soybean proteins increased more than 22 and 17%, respectively. Day and Swanson (2013) described that high chewiness in meat analogs corre- sponded with low SME values, indicative of low-melt viscosity. To summarize the literature, common soy meat analogs available in the market are presented in Table 5, which were both prominently reduced due to the dena- turation of protein molecules. Anti-nutritional factors A wide variety of anti-nutritional substances are available in most of the potential and alternative plant-derived nutrient sources. Metabolic products arising in living systems may be defined as anti-nutrient substances that affect health or food production by themselves or through their food utilization. Anti-nutritional substances could be usually divided into four groups: (1) factors that affect mineral utilization, including gossypol pigments, phy- tates (Hexa-phosphates of Myo-inositol), oxalates and glucosinolates, (2) factors that affect protein utilization and digestion, including lectins, protease inhibitors and tannins, (3) Anti-vitamins, (4) Miscellaneous substances for example, mycotoxins, cyanogens, mimosine, alka- loids, nitrate, phytoestrogens, photosensitizing agents, and saponins. Besides, Ma et al. (2020) recounted those anti-nutri- tional compounds that reduced the bio-availability of the essential nutrients or energy in the diet. Reducing the content of the anti-nutritional factors (ANFs) can efficiently improve the use of soy nutrients. Based on protein content, products from soy-protein are divided into three broad categories: (1) soy flour and grits (50% protein on a moisture-free basis); (2) soy protein con- centrates (70% protein), and (3) isolated soy proteins (90% protein) reported by Singh et al. (2008). In addi- tion, Thadavathi et  al. (2019) reported that the high proteins, and each of them has unusual denaturation temperatures. Glycinin and β-conglycinin are two main storage proteins, and their denaturation temperatures were 68 and 86°C, correspondingly (Peng et al., 2016). Throughout the low moisture extrusion, the ingredients can undergo structural changes caused by high tempera- ture and shear. It can also affect the product’s character- istics, such as microstructure and expansion (Beck et al., 2018). DSC, TG and DTG measurements The differential scanning calorimetry (DSC) analysis can be conducted with Thermal-Analysis-Systems Model Q-200. Non-isothermal degradation can be measured by using thermogravimetric/differential-thermal-analy- sis (TG/DTA). The thermal analysis techniques such as thermo- gravimetric-analysis (TGA) can provide infor- mation on thermal stability, including thermal-degra- dation of protein films. Zhang et al. (2020b) observed that the mixtures pass through extruder from the mix- ing-zone to the melting-zone for peanut protein/exoge- nous polysaccharide mixture, the endothermic-peaks of arachin and conarachin were both prominently reduced due to the denaturation of protein-molecules. In the mixing zone, 2% WS (wheat starch) could cause a significant decrease in the thermal transition peak temperature (Tp) value and a significant increase in the enthalpy changes (ΔH) value of the conarachin. As a result, it indicated that 2% WS accelerated the thermal transition of conarachin and the energy required to open the increased molecule. Guo et al. (2012) described that exogenous polysaccharides had no significant effect on the ΔH value of arachin, due to the relatively tight struc- ture of arachin. In addition, Zhang et al. (2020b) found that the exogenous polysaccharides had enhanced the protein-lipid interaction, when 0.1% CA or 2% WS was added. Moreover, the exogenous polysaccharides could decrease, particularly when 2% WS was added, and the value significantly reduced from 310 to 302°C. Textural properties There are two textural properties as the transverse (T) and directions cutting force in longitudinal (L) are pos- itively correlated with the carrageenan (ICGN) concen- tration in the meat analog sample. The cutting force can also be interpreted as an indirect indicator of texturiza- tion and hardness (Palanisamy et al., 2018). The elastic- ity differences between the raw samples containing 0.75, 1.5, and 3% ICGN were not significant, while no signif- icant differences between all ICGN added cooked sam- ples could be detected. Shahiri Tabarestani and Mazaheri Italian Journal of Food Science, 2022; 34 (2) 15 Effect of high-moisture extrusion protein vegetable-based foods were used as an alterna- tive for meat products which was enormously advanta- geous for the increasing human population in the world, and it could address the environmental concerns as well as the health considerations from meat consumption. Moreover, Aijie et al. (2014) reported that the ANFs decreased during germination, which was attributed to the activation of several enzymes in the seed. The sum- marized literature on anti-nutrients and ANFs of soy- bean are presented in Table 6. Allergenic protein There are many kinds of allergenic compounds and ANF’s present in soybean. According to the report of food allergens, almost 2% of adults and 5–8% of chil- dren have food allergic reactions associated with the consumption of soybean or soybean derived food prod- ucts (John et al., 2017). Additionally, Gagnon et al. (2010) reported that food allergies are a big concern for health in most countries. In addition, allergic reactions to food affect 4–6% of children and 1.5% of adults (Uguz et al., 2005). There are different allergenic proteins present in soybeans. However, soybean proteins aller- gic reactions are mostly transient and non-life-threat- ening and are usually outgrown after 3 years of age. It seems to be tolerant within 2–5 years after the initial diagnosis. Soybean allergenic proteins can be detecting to soybean allergens. In recent years, researches about allergenic soybean proteins detection have been rap- idly expanded. Currently using two most approaches Table 5. Common soybean meat analogs existing in market. Name of product Introduction/first reported Main ingredients/origin Characteristics/remarks References Tofu China Pressed soy curd prepared from coagulated soy. Most widely recognized meat alternatives, blind taste, can impart flavor by smoking/marinating. (Sadler, 2004) Tivall 1997 Soy-based fibrous vegetable protein. Simulate meat muscle, provide a different eating textures to other soy formats. (Sadler, 2004) Tempeh 1851 in Indonesia Fermented soy-based cake. Controlled fermentation of soy leads, similar shape to burger patties. (Kumar et al., 2017; Malav et al., 2015) Grillers original burgers Extruded vegetable protein burgers. Veggie goodness, soy protein concentrate, water for hydration. (Kyriakopoulou et al., 2019) Schnitze Rehydrated soy protein products. – (Kyriakopoulou et al., 2019) Table 6. Comparison of the soybean and other grains substitute of anti-nutritional factors and fractions. Plant substitutes Anti-nutritional factors References Soybean meal Protease inhibitors, saponins, anti-vitamins , phytic acid, lectins, phytoestrogens and allergens. (Francis et al., 2001)Rapeseed meal Rapeseed meal, protease-inhibitors, phytic acid glucosinolates and tannins. Pea seed meal Pea seed protease-inhibitors, tannins, lectins, cyanogens and phytic acid. Sesame meal Sesame phytic acid and protease inhibitors. Cottonseed meal Cottonseed meal phytic-acid, phyto-estrogens, gossypol and anti-vitamins. Various grains Grains fractions Arabinoxylans β-Glucans Cellulose Soybean Soluble – – – (Choct, 1997) Insoluble – – 4.4% Wheat Soluble 1.8% 0.4% – Insoluble 6.3% 0.4% 2% Barley Soluble 0.8% 3.6% – Insoluble 7.1% 0.7% 3.9% Maize Soluble 0.1% – – Insoluble 5.1% – 2% Sorghum Soluble 0.1% 0.1% – Insoluble 2% 0.1% 2.2% 16 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. additives, such as starch or lipid, probably have a pos- itive or negative effect on forming the desired texture. Furthermore, Asgar et al. (2010) reported that other types of legumes and oilseeds could also be used as pro- tein-rich materials to develop a variety of high-moisture meat analogs, which could contribute to alternative protein sources. Market potential alternate plant proteins In contemporary years, plant-based proteins have received increasing attention as good substitutes for animal-based proteins. The important reasons for the increasing acceptability of plant-based protein are the low cost and fibrous texture (Echeverria-Jaramillo et al., 2021). Plant-based protein’s growing trend is setting out to increase the number of vegetarians or meat avoiders (Aschemann-Witzel et al., 2021). Proteins, in particu- lar, plant-based proteins are more important in the face of future challenges, ensuing from unceasing popula- tion growth and the unevenness between malnutrition and overweight/obesity (Mittermeier-Kleßinger et al., 2021). Time trends for the development of alternative protein ingredients are demonstrated in Table 7. The key drivers of market growth include: (1) consumers con- cern over food safety in relation to animal products; (2) growth in the number of vegetarians, meat avoiders, and meat reducers; (3) meat eaters seeking more variety in their diet; (4) growing interest in healthy eating which includes incorporating more plant-based foods into the diet. Conclusion This review concluded that high-pressure extrusion is an effective technology that can potentially be used to pro- duce natural food products in which heat treatment will be reduced. This enhances the hydrophobic interactions and increases the visible viscosity to stabilize the newly formed conformation of the texturized effects. Plant proteins have some physiologically active components such as protease phytosterols, inhibitors saponins, and isoflavones. The thermo-mechanical treatment affects the microstructural changes and the rheological prop- erties in SPC during high-moisture extrusion process- ing. Therefore, the importance of soybean meat analog products as an alternative soy meat source that should be explored along with studies to clarify the underly- ing effects of the quality and increasing acceptability of plant-based protein are the low cost and fibrous texture. Future study are required on protein-extraction yield and purification which could be predicted well using another mechanistic model developed as well as on flavor and texture meat analog products. like Enzyme linked immunosorbent assays (ELISA) and Immunoblotting, also using by Radio allegro-sorbent test inhibition, Enzyme allego-sorbent test, Polymerase chain reaction, Mass spectrometry, High-performance liquid chromatography (Wang et al., 2014). ELISA is a powerful tool for detecting proteins, ELISA such as Sandwich ELISA, Competitive ELISA and Indirect ELISA are widely used to determine soybean proteins in food products (Wang et al., 2014). Immunoblotting is a powerful research tool which can indicate molec- ular mass and immunoreactivity of allergenic proteins. Beardslee (2000) found soybean allergenic proteins gly- cinin G1 acidic chain and a 22 kDa G2 glycinin by using immunoblotting. Sensory evaluation The mouthfeel and fibrousness are the same as meat, which are the primary sensory properties to consider when consuming the meat alternatives. The addition of IoT-carrageenan (ICGN) concentration is essential to increase the fibrousness of the product because of its denser structure. Palanisamy et al. (2018) observed extra 2.25 to 3% ICGN can increase elasticity which could be detected significantly with the overall scores’ acceptance ranged from 1.73 to 2.49%. The ICGN concentration was positively correlated with the fibrousness; the hardness could also be influenced by the preference (Palanisamy et  al., 2018). Cheftel et al. (1992) reported that textur- ization with HME is entirely different from other protein texturization processes such as manufacturing cheese curds, sausages, tofu, in which fiber was formed by extru- sion cooking or by spinning. In addition, proteins would be plasticized in the heating chamber during the extru- sion process of texturizing a long cooling die at the end. The process could be optimized by varying the tempera- ture, moisture, pressure, and shear. Health characteristics of soybean Regular consumption of soy products can reduce the risk of chronic diseases such as cancer, stroke, heart dis- ease, and type 2 diabetes (Jooyandeh, 2011). Soy-based foods also provide beneficial health compounds, includ- ing vitamins, minerals, fiber, and flavonoids. In addition, various clinical trials have investigated the potential of soybean and soybean products to protect against the risk of chronic diseases. Furthermore, Scheiber et al. (2001) described soybean consumption as reducing the risk of cardiovascular disease. Recently, pea protein is used as an alternative ingredient in high-moisture meat analogs because of its functional characteristics and low potentiality for allergic responses (Osen et al., 2014). Akdogan (1999) reported that protein and other food Italian Journal of Food Science, 2022; 34 (2) 17 Effect of high-moisture extrusion perspective: markets, consumers, and the challenges and oppor- tunities in the future. Crit Rev Food Sci Nutr. 61(18): 3119–3128. https://doi.org/10.1080/10408398.2020.1793730 Asgar M., Fazilah A., Huda N., Bhat R. and Karim, A. 2010. Nonmeat protein alternatives as meat extenders and meat ana- logs. Comprehen Rev Food Sci Food Saf. 9(5): 513–529. https:// doi.org/10.1111/j.1541-4337.2010.00124.x Barzegar F., Kamankesh M. and Mohammadi A. 2019. Heterocyclic aromatic amines in cooked food: a review on formation, health risk-toxicology and their analytical techniques. Food Chem. 280: 240–254. https://doi.org/10.1016/j.foodchem.2018 Beardslee T.A. 2000. IgE epitope mapping of soybean glycinin G1 acidic chain. The University of Nebraska, Lincoln. 9976975. Beck S.M., Knoerzer K., Foerster M., Mayo S., Philipp C. and Arcot  J. 2018. Low moisture extrusion of pea protein and pea fibre fortified rice starch blends. J Food Eng. 231: 61–71. https:// doi.org/10.1016/j.jfoodeng.2018.03.004 Berghout J., Pelgrom P., Schutyser M., Boom R. and Van Der Goot A. 2015. Sustainability assessment of oilseed fractionation processes: a case study on lupin seeds. J Food Eng. 150: 117–124. https://doi.org/10.1016/j.jfoodeng.2014.11.005 Bryant C. and Barnett, J. 2018. Consumer acceptance of cultured meat: a systematic review. Meat Sci. 143: 8–17. https://doi. org/10.1016/j.meatsci.2018.04.008 Caillard R., Remondetto G. and Subirade M. 2010. Rheological investigation of soy protein hydrogels induced by Maillard- type reaction. Food Hydrocolloids. 24(1): 81–87. https://doi. org/10.1016/j.foodhyd.2009.08.009 Cheftel J., Kitagawa M. and Queguiner C. 1992. New pro- tein texturization processes by extrusion cooking at high moisture levels. Food Rev Int. 8(2): 235–275. https://doi. org/10.1080/87559129209540940 Chen F.L., Wei Y.M. and Zhang B. 2011. Chemical cross-linking and molecular aggregation of soybean protein during extrusion cook- ing at low and high moisture content. LWT Food Sci Technol. 44(4): 957–962. https://doi.org/10.1016/j.lwt.2010.12.008 Chen F.L., Wei Y.M., Zhang B. and Ojokoh, A.O. 2010. System parameters and product properties response of soybean protein extruded at wide moisture range. J Food Eng. 96(2): 208–213. https://doi.org/10.1016/j.jfoodeng.2009.07.014 Choct M. 1997. Feed non-starch polysaccharides: chemical struc- tures and nutritional significance. Feed Milling Int. 191(June issue): 13–26. Day L. and Swanson B.G. 2013. Functionality of protein-fortified extrudates. Comprehen Rev Food Sci Food Saf. 12(5): 546–564. https://doi.org/10.1111/1541-4337.12023 Debruyne I. 2006. Soy base extract: soymilk and dairy alter- natives. In: Riaz, M.N., editor. Soy applications in food. CRC Press Inc, Boca Raton, USA, pp. 111–133. https://doi. org/10.1201/9781420037951.ch7 Dekkers B.L., Boom R.M. and van der Goot, A.J. 2018. Structuring processes for meat analogues. Trends Food Sci Technol. 81: 25–36. https://doi.org/10.1016/j.tifs.2018.08.011 Dixit A.K., Antony J., Sharma N.K. and Tiwari R.K. 2011. Opportunity, Challenge and Scope of Natural Products in Medicinal Chemistry, 367–383. Table 7. Time-trends for development of alternative protein ingredients. Protein ingredients Approx. year introduced References Nuts Increased imports post 1945 Soy (Sadler, 2004) QuornTM (mycoprotein) 1984 WheatProTM (wheat protein) 1992 ArrumTM (wheat and pea protein) 1995 Fibrous vegetable protein 1997 Acknowledgement This work was supported by the China Agriculture Research System (CARS-04) and the Knowledge Innovation Program Funding of the Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences (125161015000150013), “the Science and Technology Innovation Project” of the Chinese Academy of Agricultural Sciences (CAAS- ASTIP-2020-IFST-04). The first author gratefully thanks the Chinese Scholarship Council for providing a full scholarship to study PhD at the Graduate School of Chinese Academy of Agricultural Sciences, Beijing, China. Conflicts of interest There is no conflict of interest exit to the submission of this manuscript. This manuscript is approved by all authors for publication. References Aijie L., Shouwei Y. and Li L. 2014. Structure, trypsin inhibitor activity and functional properties of germinated soybean pro- tein isolate. Int J Food Sci Technol. 49(3): 911–919. https://doi. org/10.1111/ijfs.12386 Aiking H. 2011. Future protein supply. Trends Food Sci Technol. 22(2–3): 112–120. https://doi.org/10.1016/j.tifs.2010.04.005 Aiking H., de Boer J. and Vereijken J. 2006. Sustainable protein pro- duction and consumption: pigs or peas? Vol. 45. Springer Science & Business Media, Springer, Dordrecht, The Netherlands. Akdogan H. 1999. High moisture food extrusion. Int J Food Sci Technol.  34(3): 195–207. https://doi.org/10.1046/j.1365-2621. 1999.00256.x Alexandratos N. and Bruinsma, J. 2012. World agriculture towards 2030/2050: the 2012 revision, Position Paper. AgEcon Search. St. Paul, MN, USA. https://doi.org/10.22004/ag.econ.288998 Aschemann-Witzel J., Gantriis R.F., Fraga P. and Perez-Cueto  F.J. 2021. Plant-based food and protein trend from a business https://doi.org/10.1080/10408398.2020.1793730� https://doi.org/10.1111/j.1541-4337.2010.00124.x� https://doi.org/10.1111/j.1541-4337.2010.00124.x� https://doi.org/10.1016/j.foodchem.2018� https://doi.org/10.1016/j.jfoodeng.2018.03.004� https://doi.org/10.1016/j.jfoodeng.2018.03.004� https://doi.org/10.1016/j.jfoodeng.2014.11.005� https://doi.org/10.1016/j.meatsci.2018.04.008� https://doi.org/10.1016/j.meatsci.2018.04.008� https://doi.org/10.1016/j.foodhyd.2009.08.009� https://doi.org/10.1016/j.foodhyd.2009.08.009� https://doi.org/10.1080/87559129209540940� https://doi.org/10.1080/87559129209540940� https://doi.org/10.1016/j.lwt.2010.12.008� https://doi.org/10.1016/j.jfoodeng.2009.07.014� https://doi.org/10.1111/1541-4337.12023� https://doi.org/10.1201/9781420037951.ch7� https://doi.org/10.1201/9781420037951.ch7� https://doi.org/10.1016/j.tifs.2018.08.011� https://doi.org/10.1111/ijfs.12386� https://doi.org/10.1111/ijfs.12386� https://doi.org/10.1016/j.tifs.2010.04.005� https://doi.org/10.1046/j.1365-2621.1999.00256.x� https://doi.org/10.1046/j.1365-2621.1999.00256.x� https://doi.org/10.22004/ag.econ.288998� 18 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. Jooyandeh H. 2011. Soy products as healthy and functional foods. Middle-East J Sci Res. 7(1): 71–80. Joshi V. and Kumar S. 2015. Meat analogues: plant based alterna- tives to meat products – a review. Int J Food Ferment Technol. 5(2): 107–119. https://doi.org/10.5958/2277-9396.2016.00001.5 Kader M.A., Senge M, Mojid M.A. and Nakamura K. 2017. Mulching type-induced soil moisture and temperature regimes and water use efficiency of soybean under rain-fed condition in central Japan. Int Soil Water Conserv Res. 5(4): 302–308. https://doi.org/10.1016/j.iswcr.2017.08.001 Kumar P., Chatli M., Mehta N., Singh P., Malav O. and Verma A.K. 2017. Meat analogues: health promising sustainable meat substi- tutes. Crit Rev Food Sci Nutr. 57(5): 923–932. https://doi.org/10 .1080/10408398.2014.939739 Kyriakopoulou K., Dekkers B. and van der Goot A.J. 2019. Plant-based meat analogues. In: Sustainable meat production and processing. Wageningen University & Research. Wageningen, The Netherlands, pp. 103–126. https://doi.org/10.1016/B978-0-12-814874-7.00006-7 Lakemond C.M., de Jongh H.H., Hessing M., Gruppen H. and Voragen A.G. 2000. Heat denaturation of soy glycinin: influence of pH and ionic strength on molecular structure. J Agr Food Chem. 48(6): 1991–1995. https://doi.org/10.1021/jf9908704 Lin S., Huff H. and Hsieh F. 2000. Texture and chemical characteris- tics of soy protein meat analog extruded at high moisture. J Food Sci. 65(2): 264–269. https://doi.org/10.1111/j.1365-2621.2000. tb15991.x MacDonald R.S., Pryzbyszewski J. and Hsieh F.-H. 2009. Soy pro- tein isolate extruded with high moisture retains high nutri- tional quality. J Agr Food Chem. 57(9): 3550–3555. https://doi. org/10.1021/jf803435x Malav O., Talukder S., Gokulakrishnan P. and Chand S. 2015. Meat analog: a review. Crit Rev Food Sci Nutr. 55(9): 1241–1245. https://doi.org/10.1080/10408398.2012.689381 Ma M., Zhang H., Xie Y., Yang M., Tang J., Wang P., et al. 2020. Response of nutritional and functional composition, anti- nutritional factors and antioxidant activity in germinated soy- bean under UV-B radiation. LWT. 118: 108709. https://doi. org/10.1016/j.lwt.2019.108709 Mittermeier-Kleßinger V.K., Hofmann T. and Dawid C. 2021. Mitigating off-flavors of plant-based proteins. J Agr Food Chem. 69(32): 9202–9207. https://doi.org/10.1021/acs.jafc.1c03398 Olaofe O., Adeyemi F. and Adediran G.O. 1994. Amino acid and mineral compositions and functional properties of some oilseeds. J Agr Food Chem. 42(4): 878–881. https://doi. org/10.1021/jf00040a007 Orcutt M., Mcmindes M., Chu H., Mueller I., Bater B. and Orcutt A. 2006. Textured soy protein utilization in meat and meat ana log products. In: Riaz, M.N., editor. Soy applications in food. CRC Press Inc, Boca Raton, USA, pp. 155–184. Osen R., Toelstede S., Wild F., Eisner P. and Schweiggert-Weisz U. 2014. High moisture extrusion cooking of pea protein isolates: raw material characteristics, extruder responses, and texture properties. J Food Eng. 127: 67–74. https://doi.org/10.1016/j. jfoodeng.2013.11.023 Palanisamy M., Franke K., Berger R.G., Heinz V. and Töpfl S. 2019. High moisture extrusion of lupin protein: influence of extrusion Echeverria-Jaramillo E., Kim Y.-H., Nam Y.-R., Zheng Y.-F., Cho J.Y., Hong W. S., et al. 2021. Revalorization of the cooking water (Aquafaba) from soybean varieties generated as a by-product of food manufacturing in Korea. Foods. 10(10): 2287. https://doi. org/10.3390/foods10102287 Elzerman J.E., Van Boekel M.A. and Luning P.A. 2013. Exploring meat substitutes: consumer experiences and contextual factors. Br Food J. 115(5): 700–710. https://doi.org/10.1108/00070701311331490 Fang Y., Zhang B. and Wei Y. 2014. Effects of the specific mechanical energy on the physicochemical properties of texturized soy pro- tein during high-moisture extrusion cooking. J Food Eng. 121: 32–38. https://doi.org/10.1016/j.jfoodeng.2013.08.002 Fiala N. 2008. Meeting the demand: an estimation of potential future greenhouse gas emissions from meat production. Ecol Econ. 67(3): 412–419. https://doi.org/10.1016/j.ecolecon.2007.12.021 Finnigan T.J., Wall B.T., Wilde P.J., Stephens F.B., Taylor S.L. and Freedman M.R. 2019. Mycoprotein: the future of nutritious non- meat protein, a symposium review. Curr Dev Nutr. 3(6): nzz021. https://doi.org/10.1093/cdn/nzz021 Fiorentini M., Kinchla A.J. and Nolden A.A. 2020. Role of sensory evaluation in consumer acceptance of plant-based eat analogs and meat extenders: a scoping review. Foods. 9(9): 1334. https:// doi.org/10.3390/foods9091334 Francis G., Makkar H.P. and Becker K. 2001. Antinutritional fac- tors present in plant-derived alternate fish feed ingredients and their effects in fish. Aquaculture. 199(3–4): 197–227. https://doi. org/10.1016/S0044-8486(01)00526-9 Gagnon C., Poysa V., Cober E.R. and Gleddie S. 2010. Soybean aller- gens affecting North American patients identified by 2D gels and mass spectrometry. Food Anal Methods. 3(4): 363–374. https://doi.org/10.1007/s12161-009-9090-3 Golbitz P. and Jordan J. 2006. Soyfoods: market and products. In: Riaz, M.N., editor. Soy applications in food. CRC Press Inc, Boca Raton, USA, pp. 1–21. Gu B.Y. and Ryu G.-H. 2017. Effects of moisture content and screw speed on physical properties of extruded soy protein iso- late. J Korean Soc Food Sci Nutr. 46(6): 751–758. https://doi. org/10.3746/jkfn.2017.46.6.751 Guo J., Yang X.-Q., He X.-T., Wu N.-N., Wang J.-M., Gu W., et al. 2012. Limited aggregation behavior of β-conglycinin and its ter- minating effect on glycinin aggregation during heating at pH 7.0. J Agr Food Chem. 60(14): 3782–3791. https://doi.org/10.1021/ jf300409y Guo X., Sun X., Zhang Y., Wang R. and Yan X. 2018. Interactions between soy protein hydrolyzates and wheat proteins in noo- dle making dough. Food Chem. 245: 500–507. https://doi. org/10.1016/j.foodchem.2017.10.126 He L.-D., Guo X.-N. and Zhu K.-X. 2019. Effect of soybean milk addition on the quality of frozen-cooked noodles. Food Hydrocolloids. 87: 187–193. https://doi.org/10.1016/j. foodhyd.2018.07.048 John K.M., Khan F., Luthria D.L., Garrett W. and Natarajan S. 2017. Proteomic analysis of anti-nutritional factors (ANF’s) in soybean seeds as affected by environmental and genetic fac- tors. Food Chem. 218: 321–329. https://doi.org/10.1016/j. foodchem.2016.09.072 https://doi.org/10.5958/2277-9396.2016.00001.5� https://doi.org/10.1016/j.iswcr.2017.08.001� https://doi.org/10.1080/10408398.2014.939739� https://doi.org/10.1080/10408398.2014.939739� https://doi.org/10.1016/B978-0-12-814874-7.00006-7� https://doi.org/10.1021/jf9908704� https://doi.org/10.1111/j.1365-2621.2000.tb15991.x� https://doi.org/10.1111/j.1365-2621.2000.tb15991.x� https://doi.org/10.1021/jf803435x� https://doi.org/10.1021/jf803435x� https://doi.org/10.1080/10408398.2012.689381� https://doi.org/10.1016/j.lwt.2019.108709� https://doi.org/10.1016/j.lwt.2019.108709� https://doi.org/10.1021/acs.jafc.1c03398� https://doi.org/10.1021/jf00040a007� https://doi.org/10.1021/jf00040a007� https://doi.org/10.1016/j.jfoodeng.2013.11.023� https://doi.org/10.1016/j.jfoodeng.2013.11.023� https://doi.org/10.3390/foods10102287� https://doi.org/10.3390/foods10102287� https://doi.org/10.1108/00070701311331490� https://doi.org/10.1016/j.jfoodeng.2013.08.002� https://doi.org/10.1016/j.ecolecon.2007.12.021� https://doi.org/10.1093/cdn/nzz021� https://doi.org/10.3390/foods9091334� https://doi.org/10.3390/foods9091334� https://doi.org/10.1016/S0044-8486(01)00526-9� https://doi.org/10.1016/S0044-8486(01)00526-9� https://doi.org/10.1007/s12161-009-9090-3� https://doi.org/10.3746/jkfn.2017.46.6.751� https://doi.org/10.3746/jkfn.2017.46.6.751� https://doi.org/10.1021/jf300409y� https://doi.org/10.1021/jf300409y� https://doi.org/10.1016/j.foodchem.2017.10.126� https://doi.org/10.1016/j.foodchem.2017.10.126� https://doi.org/10.1016/j.foodhyd.2018.07.048� https://doi.org/10.1016/j.foodhyd.2018.07.048� https://doi.org/10.1016/j.foodchem.2016.09.072� https://doi.org/10.1016/j.foodchem.2016.09.072� Italian Journal of Food Science, 2022; 34 (2) 19 Effect of high-moisture extrusion Sha L. and Xiong Y.L. 2020. Plant protein-based alternatives of reconstructed meat: science, technology, and challenges. Trends Food Sci Technol. 102: 51–61. https://doi.org/10.1016/j. tifs.2020.05.022 Shahiri Tabarestani H. and Mazaheri Tehrani M. 2014. Optimization of physicochemical properties of low-fat hamburger formulation using blend of soy flour, split-pea flour and wheat starch as part of fat replacer system. J Food Process Preserv. 38(1): 278–288. https://doi.org/10.1111/j.1745-4549.2012.00774.x Sharma S., Thind S.S. and Kaur A. 2015. In vitro meat production system: why and how? J Food Sci Technol. 52(12): 7599–7607. https://doi.org/10.1007/s13197-015-1972-3 Shen S.H., Shi X.D. and Yuan Y.C. 2012. Optimal design of conju- gate cam-linkage combined mechanism for pressure device in die-cutting. Paper presented at the Applied Mechanics and Materials. 120: 178–181. https://doi.org/10.4028/www.scien- tific.net/AMM.120.178 Shih M.-C., Hwang T.-S. and Chou, H.-Y. 2016. Physicochemical and functional property changes in soy protein isolates stored under high relative humidity and temperature. J Food Sci Technol. 53(1): 902–908. https://doi.org/10.1007/s13197-015-2057-z Simmons A.L., Smith K.B. and Vodovotz Y. 2012. Soy ingredients stabilize bread dough during frozen storage. J Cereal Sci. 56(2): 232–238. https://doi.org/10.1016/j.jcs.2012.05.007 Singh, G. 2010. The soybean: botany, production and uses. CABI. Cambridge, MA, USA. Singh P., Kumar R., Sabapathy S. and Bawa A. 2008. Functional and edible uses of soy protein products. Comprehen Rev Food Sci Food Saf. 7(1): 14–28. https://doi. org/10.1111/j.1541-4337.2007.00025.x Singha, P. and Muthukumarappan K. 2017. Effects of processing conditions on the system parameters during single screw extru- sion of blend containing apple pomace. J Food Process Eng. 40(4): e12513. https://doi.org/10.1111/jfpe.12513 Smetana S., Mathys A., Knoch A. and Heinz, V. 2015. Meat alterna- tives: life cycle assessment of most known meat substitutes. Int J Life Cycle Assess. 20(9): 1254–1267. https://doi.org/10.1007/ s11367-015-0931-6 Smith G., Marshall W., Carpenter Z., Branson R. and Mnnke, W. 1976. Textured soy proteins for use in blended ground beef patties. J Food Sci. 41(5): 1148–1152. https://doi.org/10.1111/ j.1365-2621.1976.tb14405.x Thadavathi Y.L., Wassén S. and Kádár R. 2019. In-line rheological and microstroctural characterization of high moisture content protein vegetable mixtures in single screw extrusion. J Food Eng. 245: 112–123. https://doi.org/10.1016/j.jfoodeng.2018.10.006 Uguz A., Lack G., Pumphrey R., Ewan P., Warner J., Dick J., et al.. 2005. Allergic reactions in the community: a questionnaire survey of members of the anaphylaxis campaign. Clin Exp Allergy. 35(6): 746–750. https://doi.org/10.1111/j.1365-2222.2005.02257.x van der Weele C., Feindt P., van der Goot A.J., van Mierlo B. and van Boekel, M. 2019. Meat alternatives: an integrative comparison. Trends Food Sci Technol. 88: 505–512. https://doi.org/10.1016/j. tifs.2019.04.018 Wang F., Meng J., Sun L., Weng Z., Fang Y., Tang X., et al. 2020. Study on the tofu quality evaluation method and the establishment of parameters on extruder responses and product properties. J Sci Food Agr. 99(5): 2175–2185. https://doi.org/10.1002/jsfa.9410 Palanisamy M., Töpfl S., Aganovic K. and Berger R.G. 2018. Influence of iota carrageenan addition on the properties of soy protein meat analogues. LWT. 87: 546–552. https://doi. org/10.1016/j.lwt.2017.09.029 Paul A.A., Kumar S., Kumar V. and Sharma R. 2019. Milk ana- log: plant based alternatives to conventional milk, production, potential and health concerns. Crit Rev Food Sci Nutr. 60(18): 3005–3023. https://doi.org/10.1080/10408398.2019.1674243 Peluso I., Romanelli L. and Palmery M. 2014. Interactions between prebiotics, probiotics, polyunsaturated fatty acids and polyphe- nols: diet or supplementation for metabolic syndrome preven- tion? Int J Food Sci Nutr. 65(3): 259–267. https://doi.org/10.310 9/09637486.2014.880670 Peng X., Ren C. and Guo S. 2016. Particle formation and gelation of soymilk: effect of heat. Trends Food Sci Technol. 54: 138–147. https://doi.org/10.1016/j.tifs.2016.06.005 Pietsch V.L., Emin M.A. and Schuchmann H.P. 2017. Process con- ditions influencing wheat gluten polymerization during high moisture extrusion of meat analog products. J Food Eng. 198: 28–35. https://doi.org/10.1016/j.jfoodeng.2016.10.027 Post M.J. 2012. Cultured meat from stem cells: challenges and prospects. Meat Sci. 92(3): 297–301. https://doi.org/10.1016/j. meatsci.2012.04.008 Preece K.E., Hooshyar N., Krijgsman A., Fryer P.J. and Zuidam N.J. 2017. Intensified soy protein extraction by ultrasound. Chem Eng Process Process Inten. 113: 94–101. https://doi. org/10.1016/j.cep.2016.09.003 Riascos J.J., Weissinger A.K., Weissinger S.M. and Burks A.W. 2010. Hypoallergenic legume crops and food allergy: factors affecting feasibility and risk. J Agr Food Chem. 58(1): 20–27. https://doi. org/10.1021/jf902526y Riaz M. 2004. Texturized soy protein as an ingredient. In: Riaz, M.N., editor. Proteins in food processing. pp. 517–558. Ritala A., Häkkinen S.T., Toivari M. and Wiebe M.G. 2017. Single cell protein—state-of-the-art, industrial landscape and patents 2001–2016. Front Microbiol. 82009. https://doi.org/10.3389/ fmicb.2017.02009 Sadler M.J. 2004. Meat alternatives—market developments and health benefits. Trends Food Sci Technol. 15(5): 250–260. https://doi.org/10.1016/j.tifs.2003.09.003 Salgado J.M. and Donado-Pestana C.M. 2011. Soy as a func- tional food. In: El-Shemy H., editor. Soybean and nutrition. IntechOpen. London, UK, pp. 21–44. Samard S., Gu B.Y. and Ryu G.H. 2019. Effects of extrusion types, screw speed and addition of wheat gluten on physicochemi- cal characteristics and cooking stability of meat analogues. J Sci Food Agric. 99(11): 4922–4931. https://doi.org/10.1002/ jsfa.9722 Scheiber M.D., Liu J.H., Subbiah M., Rebar R.W. and Setchell K.D. 2001. Dietary inclusion of whole soy foods results in significant reductions in clinical risk factors for osteo- porosis and cardiovascular disease in normal postmeno- pausal women. Menopause. 8(5): 384–392. https://doi. org/10.1097/00042192-200109000-00015 https://doi.org/10.1016/j.tifs.2020.05.022� https://doi.org/10.1016/j.tifs.2020.05.022� https://doi.org/10.1111/j.1745-4549.2012.00774.x� https://doi.org/10.1007/s13197-015-1972-3� https://doi.org/10.4028/www.scientific.net/AMM.120.178� https://doi.org/10.4028/www.scientific.net/AMM.120.178� https://doi.org/10.1007/s13197-015-2057-z� https://doi.org/10.1016/j.jcs.2012.05.007� https://doi.org/10.1111/j.1541-4337.2007.00025.x� https://doi.org/10.1111/j.1541-4337.2007.00025.x� https://doi.org/10.1111/jfpe.12513� https://doi.org/10.1007/s11367-015-0931-6� https://doi.org/10.1007/s11367-015-0931-6� https://doi.org/10.1111/j.1365-2621.1976.tb14405.x� https://doi.org/10.1111/j.1365-2621.1976.tb14405.x� https://doi.org/10.1016/j.jfoodeng.2018.10.006� https://doi.org/10.1111/j.1365-2222.2005.02257.x� https://doi.org/10.1016/j.tifs.2019.04.018� https://doi.org/10.1016/j.tifs.2019.04.018� https://doi.org/10.1002/jsfa.9410� https://doi.org/10.1016/j.lwt.2017.09.029� https://doi.org/10.1016/j.lwt.2017.09.029� https://doi.org/10.1080/10408398.2019.1674243� https://doi.org/10.3109/09637486.2014.880670� https://doi.org/10.3109/09637486.2014.880670� https://doi.org/10.1016/j.tifs.2016.06.005� https://doi.org/10.1016/j.jfoodeng.2016.10.027� https://doi.org/10.1016/j.meatsci.2012.04.008� https://doi.org/10.1016/j.meatsci.2012.04.008� https://doi.org/10.1016/j.cep.2016.09.003� https://doi.org/10.1016/j.cep.2016.09.003� https://doi.org/10.1021/jf902526y� https://doi.org/10.1021/jf902526y� https://doi.org/10.3389/fmicb.2017.02009� https://doi.org/10.3389/fmicb.2017.02009� https://doi.org/10.1016/j.tifs.2003.09.003� https://doi.org/10.1002/jsfa.9722� https://doi.org/10.1002/jsfa.9722� https://doi.org/10.1097/00042192-200109000-00015� https://doi.org/10.1097/00042192-200109000-00015� 20 Italian Journal of Food Science, 2022; 34 (2) Islam M et al. analogues with hemp and soy protein using extrusion cooking. Foods. 9(6): 772. https://doi.org/10.3390/foods9060772 Zhang J., Liu L., Jiang Y., Faisal S. and Wang Q. 2020. A new insight into the high-moisture extrusion process of peanut protein: from the aspect of the orders and amount of energy input. J Food Eng. 264: 109668. https://doi.org/10.1016/j.jfoodeng.2019.07.015 Zhang J., Liu L., Jiang Y., Shah F., Xu Y. and Wang Q. 2020. High-moisture extrusion of peanut protein-/carrageenan/sodium alginate/wheat starch mixtures: effect of different exogenous polysaccharides on the process forming a fibrous structure. Food Hydrocolloids. 99: 105311. https://doi.org/10.1016/j.foodhyd.2019.105311 Zhang J., Liu L., Liu H., Yoon A., Rizvi S.S. and Wang Q. 2019. Changes in conformation and quality of vegetable protein during texturization process by extrusion. Crit Rev Food Sci Nutr. 59(20): 3267–3280. https://doi.org/10.1080/10408398.201 8.1487383 a model for suitable soybean varieties for Chinese traditional tofu processing. LWT. 117: 108441. https://doi.org/10.1016/j. lwt.2019.108441 Wang T., Qin G.-X., Sun Z.-W. and Zhao Y. 2014. Advances of research on glycinin and β-conglycinin: a review of two major soybean allergenic proteins. Crit Rev Food Sci Nutr. 54(7): 850– 862. https://doi.org/10.1080/10408398.2011.613534 Wolz M., Kastenhuber S. and Kulozik U. 2016. High moisture extrusion for microparticulation of whey proteins – influence of process parameters. J Food Eng. 185: 56–61. https://doi. org/10.1016/j.jfoodeng.2016.04.002 Wu M., Huang X., Gao F., Sun Y., Duan H. and Li, D. 2019. Dynamic mechanical properties and fractal analysis of texturized soybean protein/wheat gluten composite produced by high moisture extrusion. Int J Food Sci Technol. 54(2): 499–508. https://doi. org/10.1111/ijfs.13963 Zahari I., Ferawati F., Helstad A., Ahlström C., Östbring K., Rayner  M., et al. 2020. Development of high-moisture meat https://doi.org/10.3390/foods9060772� https://doi.org/10.1016/j.jfoodeng.2019.07.015� https://doi.org/10.1016/j.foodhyd.2019.105311� https://doi.org/10.1080/10408398.2018.1487383� https://doi.org/10.1080/10408398.2018.1487383� https://doi.org/10.1016/j.lwt.2019.108441� https://doi.org/10.1016/j.lwt.2019.108441� https://doi.org/10.1080/10408398.2011.613534� https://doi.org/10.1016/j.jfoodeng.2016.04.002� https://doi.org/10.1016/j.jfoodeng.2016.04.002� https://doi.org/10.1111/ijfs.13963� https://doi.org/10.1111/ijfs.13963�