http://isae.in/journal-of-agricultural-engineering-JAE Journal of Agricultural Engineering ISAE Vol. 58 (2): April-June, 2021 doi: 10.52151/jae2021581.1741 Production Methodologies of Meat Analogues: A Review

Girija J.1, Kamalasundari S.2, Hemalatha G.3 and Umamaheswari T.4

1P.G. Student (Food Science and Nutrition), 2Assistant Professor (Food Science and Nutrition), Department of Food Policy and Public Health Nutrition,3Professor and Head (Food Science and Nutrition), 4Assistant Professor (Agricultural Microbiology), Department of Food Science and Nutrition, Community Science College and Research Institute, Madurai. Corresponding author email address: [email protected]

Article Info ABSTRACT

Manuscript received: Meat is a non-vegetarian food and is considered as a good source of quality nutrients. March, 2020 Though meat protein provide the required content of good quality protein for the body, Revised manuscript accepted: they are also associated with higher cholesterol and fat content, which prove to be a April, 2021 leading cause of serious health issues. This became the primary reason for increase in a shift in demands for plant-based protein source foods. The other reason is environmental impact of animal derived products. Meat analogues are plant-based good quality protein source of food that tastes like meat protein, and texture resemble that of meat. These plant-based meat analogues have some amount of anti-nutrients and allergic compounds, but they can be successfully removed by employing certain processing methods and resemble meat in its functionality properties. This approach of mimicking the plant- based foods to resemble meat involves understanding of the biochemical composition and three-dimensional structure of meat, and replicating those qualities using plant-based Keywords: Plant-based ingredients. In the current scenario, the best suitable methods of manufacturing meat meat alternative, promoting analogue are by extrusion and structuring techniques. The meat analogues satisfy the vegetarianism, meat analogue need of meat for both vegetarians and non-vegetarians. This review attempts to outline manufacturing techniques, the different manufacturing processes of meat analogue using plant-based foods, and to applications analyse the best suitable method.

Meat has a distinct position in the food basket and of illness each year (Post, 2014; Grewal et al., 2014). has been widely consumed by the humans since pre- With the growing health concern of people across the historic era due to readily available source of energy, globe, the importance of health promoting foods is high quality proteins, palatability as well as images of increasing steadily that had created awareness among strength and power (Latvala et al., 2012). As a global the consumers to find meat alternatives (Aikinget al., average, per capita meat consumption has increased 2006). Animal-derived products also have challenges in to around 40 kg of meat during the year 2018 from 20 a sustainable manner. Plant-based eating is recognised kg during the year 2006. It is expected that increasing as not only nutritionally sufficient, but also as a way population, urbanization, industrialization, education to reduce the risk in all other health aspects. Meat and rise in income would cause 72 % higher demands analogues are the plant-based protein source foods that of meat consumption by the year 2030 (Godfray et al., taste like meat protein and texture resembling meat in 2010). Many researchers quote that consumption of its functionality in terms of physical properties and 120 g red meat/day or 30 g processed meat/day would sensory attributes. significantly raise the relative risk of colorectal cancer (Larsson and Wolk, 2006). The food borne pathogens Developing new meat analogues that are attractive found in meats, such as Salmonella, Campylobacter to the consumers is a challenge (Joshi et al., 2015). and E. coli are responsible for millions of episodes However, in the current scenario of technological April-June, 2021 Production Methodologies of Meat Analogues: A Review advancements, an acceptable and appreciable substitute taste identical to animal meat but are designed to be for meat products can be produced. There are several texturally similar and serve the same purpose as meat strategies to mimic the texture of meat (Narsaiah et al., in prepared foods (e.g., TVP). Since it is typically 2013), which depends on the type of meat product that used as an ingredient, functional plant-based meat may one wants to mimic. Meat products can be categorized require special formulation, just similar to how animal into ground, comminuted and whole muscle meat meat is prepared, flavoured, or tenderized for a specific products. The processed meat analogues should be able purpose. The important role of functional plant-based to mimick the original meat products. meat is replacing animal meat for reducing health claims. The main aim is to create plant-based meat from A review focussing on common forms of meat plant protein that retain their shape, texture, and flavour analogues, different protein sources used for meat through freezing, thawing, and cooking in a full range analogues, structuring and manufacturing techniques of prepared products. Certain plant and fungal foods of meat analogues from fibrous plant protein materials, can also replace animal meat as the focus of a meal by current research and future trends in meat analogue providing a whole-food product with a savoury taste processing is presented. and the mouthfeel of meat. The less processed foods, such as jackfruit, pulses, and certain mushrooms are Meat Analogue often naturally fibrous or high in protein. The list of Analogue can be defined as a compound that is ingredients used for the preparation of meat analogue structurally similar to another, but differs slightly and their purposes is given in Table 2. in composition. Meat analogue is the food which is structurally similar to meat but differs in composition. Recent researches were more focused on innovative Meat analogue, also called a meat substitute, mocks meat analogues from mushroom, pulses, and fruits meat or imitate meat approximates the aesthetic with good sensory attributes (texture, colour, flavour) qualities (primarily texture, flavour, and appearance) which is very important for acceptance of any and/or chemical characteristics of specific types of innovative product. Samard et al. (2019) compared meat (Sadler, 2004). This meat analogue is a healthier the texture characteristics of TVP among different and/or less expensive alternative to a particular meat protein products viz., isolated soy protein (ISP), isolated product due to its appearance and odour very much mung bean protein (IMBP), isolated peanut protein similar to meat (Kumar et al., 2017). Though many (IPNP), isolated pea protein (IPP), and wheat gluten vegetarian foods are available, but the variety for meat (WG) which were texturized by intermediate moisture analogue is very less. The most preferred meat analogue extrusion method. The result revealed that ISP and IPP is Texturized Vegetable Proteins (TVP). Generally, based TVPs displayed more sponge, like structure than meat analogues are made from soy-protein or gluten. the other proteins. This might be due to increasing Vegetable proteins sources are listed in Table 1. amounts of sulphur‐containing amino acids (Cystine and Methionine) of ISP, IPP and WG after extrusion. Sensory characteristics of different meat analogues The lowest essential amino acids content (Glutamic Functional plant-based meat ingredients, such as and Aspartic acid) was found in WG‐based TVP. The proteins and binding agents, are not intended to amount of loss was dependent on the intensity of

Table1. Major non-meat protein sources suitable for meat analogue

Sl . No. Type of protein Source Product Reference 1. Beta-conglycinin Soybean TVP, Tempeh, Tofu, Kinema Sun et al. (2008) 2. Glycinin, Vicilin Legumes Burmese tofu from chickpea Kang et al. (2007), Sharima et al. (2018) 3. Legumin, Albumins, Oil seeds TVP Marcone (1999); Anjum Globulins, Glutelins et al. (2011) 4. Gluten, Gliadins, Wheat, rye, barley Seitan Green and Glutenins Cellier (2007) 5. Mycoprotein Fusarium venenatum Quorn Denny et al. (Filamentous fungus) (2008)

138 Girija J, Kamalasundari S, Hemalatha G and Umamaheswari T JAE : 58 (2)

Table 2. Major Ingredients of meat analogue and their purpose

Ingredient and usage level Purpose Water - (50-80 %) Ingredient distribution, consistency of final product Textured vegetable proteins: Water binding, Soy-protein, wheat protein – (10-25 %) Texture/ mouth feel, Non texturized proteins: Appearance Egg white, whey protein- (4-20 %) Fat/ oil - (0-15 %) Flavour, texture/ mouth feel, colour Flavours/ spices - (3-10 %) Flavour: savoury, meaty, roasted, fatty, serumy, Flavour enhancement (e.g. salt) Binding agents: Wheat gluten, egg whites, gums and Texture, water binding, can determine production and hydrocolloids, enzymes, starches - (1-5 %) processing conditions Colouring agents: caramel colours, malt extracts - (0-0.5 %) Appearance/eye appeal extrusion process which determines the texture of the Maillard reaction. Wild et al., 2014 Maillard reaction meat analogue as quoted by Osen et al. (2015). occurs during heat treatment of almost every food, and bio-converts amino acids and sugars into various Anila and Suma (2018) developed TVP based on odorants, tastants and colorants as quoted by Chen et jackfruit bulb flour, seed flour and gluten in varying al. (2018). proportions. The seed flour containing treatment had high water absorption ratio as the water absorption Supplementation of flavor precursors such as amino capacity depends on the protein denaturation the acids and sugars was found to be an alternative way function of the amylase/ amylopectin ratio as well as of flavoring of meat analogue. Nivetha et al., (2019) the chain length distribution of amylopectin. Wi et al. developed meat analogue from mushroom, paneer and (2020) reported that moisture content is responsible wheat gluten flour and sensory evaluated. Among 10 for hardness, cohesiveness, springiness, and chewiness different treatments, the 50:30:20 ratio of mushroom, of meat analogue. This is also an important factor for paneer and wheat gluten flour had scored highest colour value, especially lightness of a product. Khurram overall acceptability. Mushrooms provided meaty et al. (2003) developed meat balls from isolated flavour, gluten flour served as texturizing agent, and legume-protein. The legume-proteins were isolated spices as ginger, garlic, coriander powder, pepper, chilli by adjusting the pH of the solution, precipitation, powder and salt were used for flavoring the product. coagulation and filtration. Extracted legume-protein The developed product could thus mimic the sensory isolates were incorporated with fat, and then extruded qualities of meat. to get meat-like chewiness and flavour. Although extrusion processing has been studied extensively for Manufacturing of Meat Analogue many years, the control over the process and the design Structuring techniques of extruded products is still mostly based on empirical knowledge. Producing meat in the laboratory without the involvement of living animals is a huge technical feat Besides colour and textural properties, an attractive made possible by more fundamental research into flavor is a key prerequisite for consumer acceptance stem‑cell technology and muscle development. The two and market launch of new meat analogues. A meat different structuring techniques that is generally used analogue should have a meaty flavor. These products to produce fibrous products that mimic whole muscle should be free of off-flavors, and the flavor has to meet meats are either a bottom-up or a top-down strategy to the expectations of consumers. Many meat alternatives create the fibrous morphology. developed in the past, however, lacked aroma and taste quality, which might be one reason for their Bottom-up and Top-down strategy low market acceptability. Foods can be flavored by i) In bottom-up strategy, individual structural elements addition of flavorings or spices, or ii )by odourless and are created that are subsequently assembled into tasteless ingredients that are converted during thermal larger products. This strategy follows the structural processing into sensorially active compounds as seen in organization of muscle with a hierarchy from nano-to

139 April-June, 2021 Production Methodologies of Meat Analogues: A Review macroscale, being muscle cells, myofibrils, sarcomeres substitutes are lower than beef, sometimes considerably with the proteins myosin and actin, connected via so; but tofu, novel vegetable substitutes (jackfruit) and conjunctive tissue. This arrangement follows the alternative proteins are all costlier than ground beef. structural organisation of muscle and positioning In the case of cultured beef (which has not yet reached them from nano to macroscale. The cells are grown by the market), the cost is over five times more. (World culturing or biomass production of filamentous fungi Economic Forum, 2019). or by creating protein fibrils or fibres. These fibrils/ fibres can subsequently be assembled by cross-linking Bottom-up strategy with enzymes or mixing with binding agents, such as Cultured meat egg protein and gluten to create similar structure like meat (Dekkers et al., 2018). The individual elements Cultured meat is described as ‘‘synthetic,” ‘‘in vitro,” are aligned to create similar structural anisotrophy. ‘‘artificial,” ‘‘laboratory-grown,” or ‘‘factory-grown” Cultured meat, mycoprotein, wet-spinning and electro- meat and produced via the culture of animal stem spinning use this technique. The main strength of cells into muscle cells that further develop into tissue. the bottom-up strategy is that individual structural Cultured meat is produced by tissue engineering components are assembled such that they truly resemble techniques in which in-vitro cultivation of animal the hierarchical structure of meat down to a nanoscale, cells may then be transformed into meat, instead of and thereby replicate the structure of meat in great slaughtered animals. The sensitivity of mammalian cells detail, being muscle cells, myofibrils, sarcomeres with requires a high control over the growing conditions in the protein actin and myosin connected via connective the bioreactor. To culture muscle fibres, first myoblast tissues (Post and Hocquette, 2017). cells should be harvested from the skeletal muscle from an animal. Subsequently, these myoblasts are The other technique, Top-down strategy, mimics the replicated by a standard cell culture methodology using structure on larger length scales only for which a force serum-supplemented medium with all the necessary field is used to direct structure formation or aggregation nutrients, including amino acids, lipids, vitamins and of biopolymer(s), resulting in anisotropic structure on salts, for cells to grow. When a sufficient number of a micro-meter scale . Extrusion , mixing of proteins cells are obtained, they are placed onto a scaffold with and hydrocolloids, freeze structuring and shear cell anchor points to connect and align the cells, yielding technology are done in top–down approach using this a multicellular tissue. Electric fields or other means principle. Shearing or uni-directionally freezing such are often proposed to ensure alignment and normal blends may deform the dispersed phase and align it development of muscle fibres. After three weeks, in one direction. Besides deformation, the dispersed muscle fibres get matured and can be harvested. These phase can also undergo coalesce and break-up, and fibres are 20–30 mm long and less than 1mm thick. even inversion of the two phases, which will affect the final properties of the material. After solidification, the For upscaling this technique, a large-scale reactor fibrousness in the products originates from a continuous is required in which the myoblasts can be cultured phase (mostly consisting of proteins) that is disrupted and exposed to the right environmental stimuli. by a dispersed, deformed, weak phase filaments that are The sensitivity of mammalian cells requires a high matrix/structure breakers. This affects the mechanical control over the growing conditions in the reactor and properties of the material. Therefore, the structures contamination should be avoided. These muscle fibres obtained with these techniques mimic the structure, have been used to make a single hamburger as a proof of but do not fully resemble the meat structure in its concept (Post, 2014). Processing procedure of cultured hierarchical architecture; and the question is whether the structure of the meat analogues produced with a meat production is shown in Fig.1. top-down strategy sufficiently mimics the structure of Mycoprotein real meat to entice the consumer into purchasing these products. Mycoprotein is an alternative, nutritious protein source with a meat-like texture manufactured using Fusarium The prices for the alternative proteins are not always venenatum , a naturally occurring fungus. Mycoprotein competitive with meat. The most novel alternatives based products are marketed under the brand name of are expensive, but would fall as production is scaled Quorn. Quorn products include steaks, burgers, chicken up. The costs of some established vegetable-based breasts as well as sliced meats and ready meals such as

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Cultured meat Cultured meat Farm animal

Mvofibers formation Skeletal muscle

Stem cells (mvoblast) Mvoblast proliferation Formation of Bioreactor mvotubes

Fig. 1: Processing procedure of cultured meat production

lasagna. Nutritional benefits arise from mycoprotein Waste due to its chemical composition. The cell walls of the gasses hyphae (cells) are the source of dietary fiber (chitin and glucan). The cell membranes have been reported to be the source of polyunsaturated fatty acids (PUFA’s) while the cytoplasm as the source of high quality protein. Quorn is a cholesterol free, low-fat, high-fibre meat and low cost analogue.

In UK supermarkets, a 500 g bag of frozen “Quorn” mince Ammonia Glucose and air and is approximately £2.79. A 500 g pack of ‘value’ minerals minced beef costs at least £4.00 and only increases with quality. In this method, fungus is produced Heat treatment in continuous fermentation process in bioreactors Cooling (Fig. 2). After fermentation, the RNA is degraded Drying/Chilling system into monomers by a heat treatment, so it can diffuse out of the cells. The residual biomass is heated and Mycoprotein harvested centrifuged to obtain a paste-like product with 20 % solids. The filamentous fungus is unordered after this Fig. 2: Continuous fermentation process for centrifugation step. Further process steps as forming, production of mycoprotein steaming, chilling, and texturizing, are required to obtain fibrous products. Minced-type products, such as chunks, sausages, and burgers, are commercially fibre content (6 %) imply that mycoprotein could be available from this material (Wiebe, 2004; Smetana, incorporated in various types of foods as a functional 2015). Reihani et al. (2018) produced mycoprotein ingredient (Denny et al., 2008). from date waste using Fusarium venetatum. It was observed that the use of bioreactor had a better control Wet spinning process of fermentation conditions and got higher levels of Wet spinning is mostly used for the creation of biomass, protein, and productivity. The amino and fatty individual fibres, and is one of the standard techniques acid profiles of mycoprotein, and its relatively high for the production of membranes for industrial

141 April-June, 2021 Production Methodologies of Meat Analogues: A Review separation purposes (Ho and Sirkar, 2012).Several Zein (Maize protein), when dissolved in 80 % ethanol studies showed the food-grade production of fibres with solution, are spun into fibres with a thickness in plant-based materials such as soy, pea and fababean the nanoscale (Miyoshi et al., 2005). Nieuwland et is possible. A solution containing protein is extruded al.(2014) showed the possibility to use zein as a carrier through a spinneret and subsequently immersed into for other vegetable proteins, such as soy-protein. The a bath containing a non-solvent for the exchange of proteins are required to be highly soluble and behave solvent and non-solvent resulting in precipitation and like a random coil instead of globulins. Mattice and solidification of the extruded protein phase, forming Marangoni (2020) produced fibrous material from stretched filaments with a thickness in the order of zein by using different methods (electro-spinning, anti- 20 μm. Using immersion rotary jet spinning, a dry-jet solvent precipitation and mechanical elongation) for the wet-spinning process; gelatin fibres are produced at intended use of structuring whole tissue meat analogues. high rates comparable to natural collagen fibres could The result showed that mechanical elongation displayed be developed. The type of structure that is formed the greatest potential in structuring whole-tissue meat depends on the solidification mechanism. The fibres analogues, as it produced a statistically similar texture are obtained when the dispersed phase solidifies and to chicken meat. the continuous phase can be washed away; capillary filled gels are obtained when the continuous phase is Top-down strategy solidified and the dispersed phase stays liquid, and fibre Extrusion filled gels are obtained when both the dispersed and the Extrusion is currently the best and most commonly continuous are solidified. used commercial technique to transform plant-based materials into fibrous products for production of meat MacQueen et al.(2019) developed meat products from analogues. Extrusion is relatively energy intensive gelatin fibres by wet spinning method. As gelatin and applies thermo-mechanical treatment using high is a natural component of meat, extruded gelatin temperatures and shear rates inside the barrel/screw microfibers recapitulated structural and biochemical region resulting in melting of the protein suspension features of natural muscle tissues during processing and intensive mixing. In extrusion, the proteins are and cooking. plasticized inside the barrel by a combination of heating, hydration and mechanical deformation. When Electrospinning method this protein-‘melt’ flows into the dye, it gets aligned by Electrospinning can also be used to produce fibres laminar flow, and is cooled to prevent expansion. The for the application of meat analogues (Nieuwland processing of meat analogues in such cooking extruders et al., 2014). Electrospinning produces individual involves a multitude of machine and process parameters. fibres of the smallest scale. In electrospinning, a Furthermore, the composition of the matrix, the variety biopolymer solution is pushed through a hollow needle of ingredients and the water content has a major effect or spinneret that has an electric potential relative to on the final product. According to the model developed a ground electrode. Accumulation of charge on the by Mitchell and Areas (1992), the biopolymer melts surface of the droplet that emerges from the spinneret and has two phases: (a) a homogenous continuous causes surface instabilities that ultimately grow into phase, and (b) a dispersed insoluble phase. Extrusion of very thin fibres (≈100 nm), which are attracted to materials/ingredients depend on the ratio of soluble and the ground electrode (Schiffman and Schauer, 2008). insoluble components; too many insoluble components Food-grade electrospinning is mostly presented for disturb protein cross-linking and result in incoherent applications in which nanofibres were used as carriers products. The extrusion process of meat analogue is or delivery systems for bioactive components such shown in Fig. 3. as polyphenols and probiotics (Libran et al., 2017). Electrospinning of proteins has been reported for There are two classes of extrusion methods. They several animal-based proteins such as whey, collagen, are low-moisture and high-moisture extrusion. In egg, and gelatin, but only sparingly for plant proteins low-moisture extrusion, flours or concentrates are as plant proteins are in their native state globular; and mechanically processed into texturized vegetable upon denaturation, insoluble aggregates are formed proteins (TVP), which are dry, slightly expanded (Anu and Anandharamakrishnan, 2014; Ghorani and products and are moisturized afterwards. Sun et al. Tucker, 2015). (2021) commented that meat analogues made via ‘‘dry

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Although the emulsion process made proteins more susceptible to thermal degradation, the hydrocolloids used in this research did not affect the formation of protein gel or the formation of interstitial film in the emulsions. This process is well scalable, yields products with some degree of structure, but still is relatively intensive in its use of resources.

Freeze structuring In freeze structuring or freeze alignment, an aqueous

solution or slurry of proteins is frozen to generate Fig. 3: Process of extrusion of meat analogue Fig. 3: Process of extrusion of meat analogue structure. Directional freezing has been studied for extrusion” (moisture < 30 %) have limited acceptance structuring of meat, fish, and plant proteins (Lugay and Kim, 1978; Consolacio and Jelen, 1986).The size of because of their poor mouth feel. Meanwhile, ‘‘wet extrusion” under high moisture conditions (40%– the ice crystal needles can be tailored with the freezing

80%) enables the production of fresh and premium temperature .Subsequently, the frozen product is dried meat analogues, with a muscle meat-like texture as without melting the ice crystals (for example by freeze well as a similar appearance and chewing sensation to drying) to obtain a porous microstructure with sheet cooked meat. Lin et al. (2002) reported that soy-protein like parallel orientation of the proteins. These sheets extruded under high-moisture conditions can have well- are connected forming a cohesive fibrous product defined fibre formations resembling that of chicken or (Consolacion and Jelen, 1986). To obtain distinct turkey breast meat. TVP is produced using hot extrusion fibrous products, the proteins should have relatively of defatted soya proteins, resulting in expanded high good solubility prior to freezing, and during the freezing protein chunks, nuggets, strips, grains and other shapes process these proteins become insoluble. The novel nd chickpea. The fibrous, insoluble, porous TVP can freezing technology have positive effect on promoting soak up water or other liquids a multiple of its own the formation of better microstructure, and enhanced quality attributes (Zhan et al., 2018).Heat removal weight. Textured soy-proteins (TSP) are processed to impart a structure and appearance that resembles from a well-mixed slurry leads to an isotropic structure. meat, seafood or poultry, when hydrated. Soy-protein When heat is removed unidirectionally without mixing, products have become increasingly popular because of the alignment of ice crystal needles yields anisotropic their low price, high nutritional quality, and versatile structures. functional properties (Joshi and Kumar, 2015). Shear cell technology Mixing of proteins and hydrocolloids Shear cell technology is a new technique based on Fibrous products can be obtained by mixing protein the concept of flow-induced structuring. A cone–cone with hydrocolloids that precipitate with multivalent device based on a cone–plate rheometer was developed cations (Kweldam, 2011). After mixing, the fibrous and referred as shear cell (Grabowska et al., 2016). The products are washed, and the excess water is removed top cone is stationary while the bottom cone rotates. by pressing. The residual dry matter contents are Both cones can13 be heated and cooled with the use of an between 40 % and 60 % moisture content on wet basis. oil bath. Contrary to extrusion, the deformation inside Plant proteins such as soy, rice, maize and lupine can the device is well defined and constant upon processing. be employed in a similar way. Due to a combination of simple shear and heat, proteins are aligned forming fibrous meat substitutes. Mejia et al. (2018) evaluated the physico-chemical effects of incorporation of β-glucan in a meat emulsion The final structure of the product obtained from this through optimal mixture modelling systems. The technique depends on the ingredients and on the optimal mixture design produced in this study was processing conditions. Fibrous products are obtained done with the incorporation of other hydrocolloids with calcium caseinate and several plant protein carrageenan (C) and starch (S). Texture profile analysis blends, such as soy-protein concentrate, soy-protein parameters were influenced by the presence of β glucan isolate (SPI), wheat gluten (WG) and SPI - pectin. The and starch indicating their participation in the processes structures prepared with calcium caseinate showed of moisture retention and structuring emulsions. anisotropy on a nanoscale, while for the plant-based

143 April-June, 2021 Production Methodologies of Meat Analogues: A Review material anisotropy was observed up to the micrometre- involved besides disulfide bonds. The results indicated scale. Grabowska et al. (2016) studied that shear- that disulfide bonding played a more important role induced structuring can be used to make anisotropic than non-covalent bonds in not only holding the rigid materials using a mixture of soy-protein concentrate structure of extrudates, but also forming of fibrous and water only. texture. The sharpest decrease in protein solubility occurred when the mix passed through the intermediate Schreuders et al. (2019) developed a product from pea section of the extruder barrel, indicating formation protein isolate and wheat gluten into materials with a of new disulfide bonds during the stage of dramatic fibrous morphology using shear induced structuring increase in both temperature and moisture. combined with heating. The study resulted in distinct fibrous morphology when sheared and heated at 120 °C. Rehrah et al.(2009) determined the extrusion processing Processing at a lower temperature resulted in a weak parameters for developing texturized meat analogue product without fibres, while higher temperature gave (TMA) from defatted peanut flour. The result showed a strong and layered product. In contrast, soy-protein that the extrusion conditions that produced the best isolate and wheat gluten blends yielded similar fibrous, TMA were60–65 % protein, 50–55 % moisture, anisotropic materials at a broader range of shearing extruded at 160–165 0C with screw speed of 80–90 rpm. temperatures (110–140 °C). Krintiras et al.(2015) Sensory acceptability of snacks incorporating peanut applied couette cell concept for the production of based TMAs was better than those containing soya- structured meat analogues via simple shear flow by based TMAs in terms of flavour, texture, off-flavour, using SPI and WG dispersion. SPI and gluten were and overall liking. Yadav et al.(2015) prepared meat sheared independently as well as in a mixture of both. It analogue rolls with a combination of selected cereal was found that SPI alone could not yield any structure, mix base (50 %) + textured soy-grit (soaked) 25 % whereas gluten alone at 30 % blend could form fibrous + mushroom paste 25 per cent The texture profile structures. The sheared SPI–gluten blend resulted analysis and sensory scores of prepared meat analogue in anisotropic structures and possible to structure was almost similar to chicken meat rolls. Arueya et SPI–WG blends into fibrous anisotropic and layered al.(2017) developed texturized vegetable protein (TVP) materials using a Couette Cell under realistic process from under-utilized sources (Lima bean (90 percent) conditions (temperature: 90–1100C, process time: 5–25 and African oil bean seed protein concentrate of 1 per min, rotation rate: 5–50 rpm). The result showed that cent.) by using extrusion processing conditions with the application of simple shear and heat is the key in optimum parameters of barrel temperature (92.45 0C), obtaining structured plant protein-based products. screw speed (101.48 rpm), feed moisture (59.63 %).

Current Researches on Meat Analogues Sharima-Abdullah et al.(2018) developed imitation Texture, appearance, and flavour are the three major chicken nuggets (ICN) from chickpea flour and components of food acceptability. In terms of consumer textured vegetable-protein in different combinations. preference, structuring of non-meat proteins are Results revealed that all ICN had significantly lower essential for development of meat analogue. The fibrous values (P<0.05) of cooking loss, lightness, hardness, texture of meat can be mimicked in plant-proteins by chewiness, springiness, cohesiveness, and water texturization. Meat analogue can also be mimicked with activity compared to control nugget. The most preferred latest technologies to resemble ground, communited, nugget combination was 10 % chickpea flour and and whole muscle meat products. 30 % TVP. Samard and Ryu (2019) investigated the physico-chemical properties, texture, and structure Liu and Hsieh et al. (2008) developed extruded meat (viz., nitrogen solubility and integrity index, chewiness analogues from soy-protein mixed with gluten and and cutting strength) of TVP and compared with starch under high moisture and high-temperature different types of meats (beef, pork, and chicken). The conditions. The protein solubility of samples collected result showed that the physio-chemical property like at different extruder zones and extrudates were made water absorption capacity of TVP (2173.84 g.kg-1) was by 11 extraction solutions of phosphate salts, urea, significantly higher than all meat samples (1095.37 to DTT, thiourea, Triton X-100, and CHAPS. Among 1653.52 g.kg-1). Due to the difference in protein types, these reagents, isoelectric focus (IEF) buffer solubilized the colour and amino acids of TVP were different from the highest levels and equal amounts of proteins in all meat samples. TVP showed a fibrous structure with samples indicating that there are no other covalent bonds non-uniform air cells.

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Chiang et al. (2019) investigated the physico-chemical diet as also for exploring new food products. Ethical properties of sausages made from meat alternatives reasons viz., strict environmental controls, increased that is Maillard-reacted beef bone hydrolysate and nutrient fortification, individually-customized cellular plant-proteins (soy-protein and wheat-gluten) at and molecular compositions, nutritional profiles, and different moisture contents (49- 60 %, w.b.). The tissue monitoring that prevent infections of meat study concluded that hardness and chewiness of meat cultures making it healthier than livestock-sourced analogue sausages decreased as moisture content (MC) meat also would play an important role. increased. The sample (49 % MC) had the highest sensory score among all sausages made from meat Recent researches are focussed to develop meat alternatives. Meat alternatives at even lower MC (<49 analogue from plant-protein along with animal-protein. %) was recommended to retain higher hardness when Texturized wheat gluten is commercially available in making into sausages. several forms differing in size, shape, density, colour, and texture. The popularity of texturized wheat gluten Future Trends of Meat Analogues is rapidly increasing due to abundant production of The strength of development of plant-based meat wheat throughout the globe. Researchers are attempting analogues is believed to be a great way to improve to develop wheat varieties that have higher amount human health, conserve natural resources and conserve of gluten while maintaining its shear flow (viscosity, animals. Another important reason for the increased extensibility, elasticity) properties. Genetic engineering acceptance of plant-protein is their low cost and fibrous can enhance the quality of plant-based food products texture, and a good alternative to animal food products. through silencing of genes. New plant-based meat The price offered is relatively affordable because of anlogues should taste, feel, and smell better, or at least use of plant-based food preparations, as from pulses as good as animal meat, satisfying the perceptions of or grains. These raw materials are readily available, the consumers. Probably umami flavour associated can be stored, and are affordable. Some of these plant- with meat and meat product fibre-like textures are protein preparations as tofu, tempeh, and seitan have the important keys to success, and also the biggest high protein contents, and can compete with meat or challenges for the researchers. fish both in their nutritional value and in their function as outstanding food components. Besides nutritional The meat substitutes market in India is estimated to value, many functional components help in preventing reach over $ 47.57 million in value terms by the end of and curing diseases, which means that they fall in the the year 2026, and is forecasted to grow at Compound scope of medicinal food homology (MFH). However, Annual Growth Rate (CAGR) of 7.48% during 2021- plant-based analogue products are not widely known to 2026. This market is driven by rising health concerns the consumers. Lack of awareness on the use of simple coupled with growing awareness about various diseases and appropriate production process technologies is also caused due to lack of proteins. Moreover, the market a weakness. is segmented based on product, source, type, form, manufacturer, and region (TechSci Research, 2021). The major challenging task for the food engineers, It can be concluded that there is a demand as well as however, is to develop the fibrous three-dimensional bright future of such products in the Indian market, structures from these plant proteins and by maintaining keeping aside a few constrains that need solutions, but the nutritional properties so as to provide these alternate with a heap of opportunities. meat products the same meaty texture. As the products have a good potential market for a product innovation in CONCLUSIONS the form of meat analog. Many researches have shown great interest in the possibility of fabricating protein- Human health is influenced more by food than by any rich food from plant sources in a palatable form and shift other single factor, and facilitating healthy diets is the mindset of policy makers and scientists towards the critical both for individual wellbeing and containing development of suitable alternatives of simulated meat- the costs of treating illnesses. There are different like products with controlled texture, flavour, colour, processing routes to create fibrous products, which and nutritive value. The market opportunities for meat can be classified in bottom-up and top-down approach. analogue do not only include vegetarians, but also non- The bottom-up approach starts with the formation vegetarians who seek to reduce the meat consumption of individual structural components from individual for health concerns through sustainable and convenient molecules or components, including proteins that are

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