A method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes

Abstract

This invention discloses a method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzyme, comprising the following steps: (1) mixing gluten-free dual-protein raw materials, the dual-protein raw materials including soy protein isolate and pea protein isolate; (2) TG enzyme incubation, the amount of TG enzyme added is 0.2% to 1.0% of the total dry basis; (3) high-moisture extrusion, the moisture content is 55% to 65%; (4) cooling and cutting; (5) processing of plant-based meat products. In this invention, during the extrusion process, TG enzyme-induced protein gelation and fiber structure formation occur simultaneously and compete with each other. 11S globulin in soy protein isolate is highly sensitive to TG enzyme, and rapidly forms a covalent cross-linked network during extrusion, constituting a fiber skeleton structure; pea protein isolate is relatively insensitive to TG enzyme, and its moderate gelation slows down the overall gelation rate, providing sufficient development time and space for the directional arrangement of fiber structures along the extrusion direction.

Description

Technical Field

[0001] This invention relates to the field of plant-based meat product processing, specifically to a method for preparing plant-based meat by using soy protein isolate (SPI) and pea protein isolate (PPI) as raw materials, utilizing the differences in the response characteristics of the two proteins to transglutaminase (TG enzyme), and through the competitive balance between gelation and fibrosis during high-moisture extrusion. Background Technology

[0002] High-moisture extrusion technology (material moisture content ≥40%) is one of the most efficient manufacturing technologies in the current plant-based meat processing field, producing extrudates with fibrous structures and textures closer to animal meat. TG enzymes, as enzymes that catalyze covalent cross-linking between protein molecules, work synergistically with protein raw materials during high-moisture extrusion to effectively improve the fibrous structure and textural properties of plant protein extrudates. However, the response characteristics of plant proteins from different sources to TG enzymes vary significantly. Previous studies have shown that the protein recombination catalyzed by TG enzymes during high-moisture extrusion is affected by the type and concentration of the protein. TG enzymes significantly reduce the hardness of peanut protein extrudates, significantly increase the hardness of soy protein isolate extrudates, but have no significant effect on pea protein isolate extrudates. Furthermore, protein gelation and fiber structure formation compete during high-moisture extrusion; excessively rapid gelation is detrimental to the full development of fiber structures.

[0003] Currently, there is no existing technology that utilizes the difference in the response characteristics of soy protein isolate and pea protein isolate to TG enzymes to synergistically regulate fiber structure. The purpose of this invention is to directionally prepare plant-based meat products with ideal fiber structure by rationally designing the ratio of the two proteins and regulating the balance between gelation and fibrinization during high-moisture extrusion using TG enzymes. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention discloses a method for preparing high-moisture extruded plant-based meat by utilizing the difference in transglutaminase (TG) response between soy protein isolate and pea protein isolate to regulate fiber structure. During extrusion, TG enzyme-induced protein gelation and fiber structure formation occur simultaneously and in competition. The 11S globulin in soy protein isolate is highly sensitive to TG enzyme, rapidly forming a covalently cross-linked network during extrusion to constitute the fiber backbone structure. Pea protein isolate is relatively insensitive to TG enzyme; its moderate gelation slows down the overall gelation rate, providing sufficient time and space for the directional alignment of the fiber structure along the extrusion direction. By controlling the ratio of soy protein isolate to pea protein isolate and the amount of TG enzyme added, precise control of the competitive balance between gelation and fiber formation can be achieved.

[0005] This invention is achieved through the following technical solution: A method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes includes the following steps: (1) Gluten-free dual-protein blend; (2) TG enzyme incubation; (3) High moisture content extrusion; (4) Cooling and cutting; (5) Processing of plant-based meat products.

[0006] Specifically, the following steps are included: 1. Gluten-free dual protein ingredient blend Weigh the following raw materials by weight: 50-70 parts soy protein isolate (protein content ≥90%) and 20-50 parts pea protein isolate (protein content ≥80%). Put the above protein raw materials into a high-speed mixer and mix at 600-1000 r / min for 5-10 min until homogeneous.

[0007] 2. TG enzyme incubation Add 0.2%–1.0% (dry weight) of TG enzyme to the mixed protein raw materials and continue mixing for 3–5 minutes to ensure uniform enzyme distribution. Let stand at 4°C for 12 hours. Utilizing the natural differences in the TG enzyme response capabilities of soy protein isolate and pea protein isolate, a synergistic regulatory system is formed where soy protein isolate exhibits a high TG enzyme response, forming the gel framework, while pea protein isolate exhibits a low TG enzyme response, delaying the gelation process.

[0008] 3. High moisture content extrusion The dual-protein system was fed into a twin-screw extruder via a loss-in-weight feeder for high-moisture extrusion texturing. During extrusion, water was added online to control the material's moisture content to 55%–65%. The extruder temperatures were set as follows: feeding zone 40–70℃, mixing zone 70–110℃, cooking zone 140–160℃, cooling zone 100–130℃, and forming zone 50–80℃. The screw speed was 260–300 r / min, and the feeding rate was 12–20 kg / h.

[0009] 4. Cooling and Cutting After extrusion molding, the textured protein strips are cooled to 45°C via a 5-meter-long cooling conveyor belt and then cut into 20-cm-long strips to obtain high-moisture composite textured protein.

[0010] 5. Processing of plant-based meat products Take the above-mentioned textured protein, and season it through processes such as desiccation, cutting, or tumbling to prepare corresponding plant-based meat products.

[0011] Beneficial effects Soy protein isolate exhibits a low threshold and high response, while pea protein isolate exhibits a high threshold and progressive response. This invention utilizes the inherent differences in the transglutaminase responses of soy protein isolate and pea protein isolate to form a gel system consisting of a rapid SPI gel framework and a PPI-delayed gel filling. This system achieves targeted regulation of fiber structure without relying on gluten-containing proteins (gluten powder), offering advantages such as gluten-free processing and a wide processing window. Attached Figure Description

[0012] Figure 1 This is a mechanism for balancing the gelation and fibrinization of high-moisture extruded proteins. Figure 2 This is a product image of high-moisture extruded protein. Detailed Implementation

[0013] The following embodiments and accompanying drawings are merely illustrative of specific implementation schemes for carrying out the present invention. These schemes and drawings should not be construed as limiting the present invention. Any changes made without departing from the principles and essence of the present invention shall fall within the protection scope of the present invention.

[0014] Unless otherwise specified, the experimental techniques and methods used in this embodiment are conventional. Unless otherwise specified, all materials and reagents used in this embodiment can be obtained through legitimate commercial channels. Soy protein isolate was purchased from Shandong Yuxin Biotechnology Co., Ltd.; pea protein isolate was purchased from Yantai Shuangta Food Co., Ltd.; and TG enzyme was purchased from Zhongnuo Biotechnology Development Jiangsu Co., Ltd.

[0015] Example 1 Soy protein isolate and pea protein isolate were mixed in a 50:50 ratio in a high-speed mixer at 800 r / min for 10 min until homogeneous. TG enzyme was added at 0.8% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder via a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone, with a feeding speed of 15 kg / h. The material was then extruded and shaped.

[0016] Take 1000g of extruded textured soy protein, break it into 2cm long strands, and add 90g of soybean oil, 70g of potato starch, 16g of methylcellulose, 40g of chicken flavor seasoning, and 0.2g of red yeast rice in sequence. Stir slowly for 6 minutes until well mixed. Press the mixture into patties with a diameter of 9cm and a thickness of 1.2cm (approximately 100g each), and quick-freeze and package them as chicken patty products.

[0017] The parameters were varied: the ratio of soy protein isolate to pea protein isolate was 50–70:30–50; the TG enzyme content was 0.2–0.8%; the cooking zone temperature was 140–160℃; the moisture content was 55–65%; and the screw speed was 260–280 r / min. The specific effects on the texture and organization of the textured protein are shown in Tables 1 and 2.

[0018] Example 2 The ratio of soy protein isolate to pea protein isolate was 60:40. They were mixed evenly in a high-speed mixer at 800 r / min for 10 min. TG enzyme was added at a rate of 0.8% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone. The feeding speed was 15 kg / h, and the material was extruded and shaped.

[0019] Take 1000g of extruded protein, break it into 3mm granules, add 80g of soybean oil, 60g of tapioca starch, 20g of methylcellulose, 50g of beef flavor seasoning, 0.4g of caramel color, and 0.2g of beetroot red. Mix well and press into meat patties with a diameter of 9cm and a thickness of 1.0cm. Quick-freeze and package as a beef patty product.

[0020] Hardness, elasticity, and chewiness: The testing instrument was a TA-XT plus texture analyzer. A P50 probe was used. The sample was cut into 20mm × 20mm × 7mm cubes. The specific testing parameters were set as follows: initial speed 1mm·s. -1 The test speed is 1 mm·s -1 The speed after testing was 2 mm·s. -1 The pressure was 50%, repeated twice, and each group of samples was tested 10 times. The average value was taken after removing the two maximum and two minimum values ​​for significance analysis.

[0021] Cooking loss rate: The weight difference method was used to accurately weigh the plant-based meat sample before cooking (W1); the sample was placed in a sealed container and heated in boiling water at 100℃ until the center reached 75℃, and maintained for 20 minutes; the sample was removed, cooled to room temperature, and the surface moisture was dried with absorbent paper, and then weighed again (W2); the cooking loss rate (%) was calculated as (W1-W2) / W1×100%.

[0022] Method for determining texture: The testing instrument was a TA-XT plus texture analyzer. An HDP / BS probe was used. Samples were cut into 20mm × 20mm × 7mm cubes, and the texture of the plant protein meat was determined using the Warner-Bratzler shearing method. Specific parameters were set as follows: pre-test speed was 1 mm / s. -1 The test speed is 1 mm·s -1 The speed after testing was 2 mm·s. -1 The degree of shearing was 75%. Shear forces in the vertical and parallel directions were measured separately. The ratio of vertical shear force to parallel shear force was defined as the degree of tissue formation. Each group of samples was tested 10 times. The two maximum and two minimum values ​​were removed and the average value was used for significance analysis.

[0023] Table 1. Effects of extrusion parameters on the texturization of SPI extruded fibroin Table 2. Effects of TG enzyme on the structure of proteins extruded and drawn into fibers at different ratios. Table 3. Cooking test indicators for chicken patties and beef patties Comparative Example 1 Soy protein isolate and pea protein isolate were mixed in a 50:50 ratio in a high-speed mixer at 800 r / min for 10 min until homogeneous. TG enzyme was added at 0.1% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone. The feeding speed was 15 kg / h, and the material was extruded and shaped.

[0024] Comparative Example 2 Soy protein isolate and pea protein isolate were mixed in a 50:50 ratio in a high-speed mixer at 800 r / min for 10 min until homogeneous. TG enzyme was added at 1.2% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone. The feeding speed was 15 kg / h, and the material was extruded and shaped.

[0025] Comparative Example 3 The TG enzyme content is 0.2% of the total mass of soybean protein isolate raw material. After standing at 4℃ for 12 hours, the protein system is fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion and texturization treatment. During the extrusion process, the material moisture content is 60%, the screw speed is 280 r / min, the temperature of the feeding zone is 50℃, the temperature of the mixing zone is 100℃, the temperature of the cooking zone is 150℃, the temperature of the cooling zone is 120℃, the temperature of the forming zone is 60℃, the feeding speed is 15 kg / h, and the extrusion is shaped.

[0026] Comparative Example 4 The TG enzyme content is 0.2% of the total mass of pea protein isolate raw material. After standing at 4℃ for 12 hours, the protein system is fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion texturization. During the extrusion process, the material moisture content is 60%, the screw speed is 280 r / min, the temperature of the feeding zone is 50℃, the temperature of the mixing zone is 100℃, the temperature of the cooking zone is 150℃, the temperature of the cooling zone is 120℃, the temperature of the forming zone is 60℃, the feeding speed is 15 kg / h, and the extrusion is shaped.

[0027] Comparative Example 5 Soy protein isolate and peanut protein isolate were mixed in a 50:50 ratio in a high-speed mixer at 800 r / min for 10 min until homogeneous. TG enzyme was added at 0.2% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder through a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone, with a feeding speed of 15 kg / h. The material was then extruded and shaped.

[0028] Comparative Example 6 Pea protein and peanut protein isolate were mixed in a 50:50 ratio in a high-speed mixer at 800 r / min for 10 min until homogeneous. TG enzyme was added at a rate of 0.2% of the total mass of the mixed protein raw materials. After standing at 4℃ for 12 h, the dual-protein system was fed into a twin-screw extruder via a loss-in-weight feeder for high-moisture extrusion texturization. During extrusion, the material moisture content was 60%, the screw speed was 280 r / min, the temperature was 50℃ in the feeding zone, 100℃ in the mixing zone, 150℃ in the cooking zone, 120℃ in the cooling zone, and 60℃ in the forming zone, with a feeding speed of 15 kg / h. The material was then extruded and shaped.

[0029] The extruded textured protein prepared in Comparative Examples 1-6 was subjected to texturing and textural tests, as detailed in Table 4.

[0030] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes, characterized in that, Includes the following steps: (1) Gluten-free dual-protein blend, the dual-protein blend including soy protein isolate and pea protein isolate; (2) Incubation with TG enzyme, the amount of TG enzyme added is 0.2% to 1.0% of the total dry weight; (3) High moisture content extrusion, with a moisture content of 55% to 65%; (4) Cooling and cutting; (5) Processing of plant-based meat products.

2. The method for preparing high-moisture extruded plant-based meat based on TG enzyme differential regulation according to claim 1, characterized in that, The mass ratio of soy protein isolate to pea protein isolate is 50–70:30–50.

3. The method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes according to claim 1, characterized in that, Mix the gluten-free dual protein ingredients at 600–1000 r / min for 5–10 min until homogeneous.

4. The method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes according to claim 1, characterized in that, The amount of TG enzyme added is 0.2% to 0.8% of the total dry weight, and the incubation time is 12 h at 4℃.

5. The method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes according to claim 1, characterized in that, The temperatures in each zone during the extrusion process are set as follows: feeding zone 40-70℃, mixing zone 70-110℃, cooking zone 140-160℃, cooling zone 100-130℃, forming zone 50-80℃, screw speed 260-300 r / min, and feeding speed 12-20 kg / h.

6. The method for preparing high-moisture extruded plant-based meat based on differential regulation of TG enzymes according to claim 5, characterized in that, During the extrusion process, the screw speed is 280 r / min, the temperature of the feeding zone is 50℃, the temperature of the mixing zone is 100℃, the temperature of the cooking zone is 150℃, the temperature of the cooling zone is 120℃, the temperature of the forming zone is 60℃, and the feeding speed is 15 kg / h.

Images