Vegetable protein composition and production method thereof

Abstract

The invention provides a vegetable protein composition and a production method thereof, and belongs to the technical field of protein. Comprising the following steps: S1, removing bean substance resistance factors; s2, carrying out mixed fermentation on the microalgae, bean substances and protein mulberry, and separating to obtain protein peptide; s3, chelating metal ions of the protein peptide; s4, condensing the protein peptide chelated with the metal ions; s5, preparation of carboxylated pullulan polysaccharide; s6, preparation of hydrophobic modified carboxylated pullulan polysaccharide; and S7, preparing the plant protein composition. The prepared plant protein composition has good physiological activity, through the embedding technology, the storage stability, heat resistance, acid resistance and moisture resistance of protein peptide are improved, growth and development of livestock are promoted, the meat quality is improved, and the plant protein composition has wide application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of proteins, and in particular to a plant protein composition and a production method thereof. Background Art

[0002] Feed is the material foundation for the development of animal husbandry, and protein feed is a crucial component. Feed raw materials are generally rich in protein, which typically requires proteases to break down into peptides and amino acids for reuse. Many probiotics, such as Bacillus, Aspergillus niger, and Aspergillus oryzae, secrete proteases that break down and utilize protein in the environment, thereby promoting their own growth. However, Saccharomyces cerevisiae and lactic acid bacteria secrete fewer proteases and have a lower ability to degrade protein materials. They generally require supplementation with protein-degrading metabolites such as peptone for optimal growth. Acidic or neutral proteases are also commonly used in feed. Proteases are categorized as endo- and exo-enzymes based on their mode of action. Microbial proteases are typically found as mixtures of endo- and exo-enzymes. Most animals require protease supplementation. For example, early-weaned piglets often require protease supplementation to compensate for insufficient digestive enzymes.

[0003] Plant protein raw materials are widely available and inexpensive, making them an ideal fishmeal substitute. Research has shown that partially or completely replacing protein in feed with plant protein does not affect animal growth performance, demonstrating their significant research significance and broad application potential. However, plant protein raw materials also have certain drawbacks, such as the presence of various anti-nutritional factors, amino acid imbalances, and high cellulose content. Exceeding a certain level of fishmeal replacement can negatively impact the growth and health of aquatic animals. Summary of the Invention

[0004] The purpose of the present invention is to provide a plant protein composition and a production method thereof, which has good physiological activity. Through encapsulation technology, the storage stability, heat resistance, acid resistance and moisture resistance of the protein peptide are improved, the growth and development of poultry and livestock are promoted, the meat quality is improved, and the application prospects are broad.

[0005] The technical solution of the present invention is achieved as follows:

[0006] The present invention provides a method for producing a plant protein composition, characterized in that it comprises the following steps:

[0007] S1. Removal of resistance factors in legumes;

[0008] S2. Fermentation of microalgae with legumes and protein mulberry to separate and obtain protein peptides;

[0009] S3. Metal ion chelation by protein peptides;

[0010] S4. Condensation of protein peptides that chelate metal ions;

[0011] S5. Preparation of carboxylated pullulan;

[0012] S6. Preparation of hydrophobically modified carboxylated pullulan;

[0013] S7. Preparation of plant protein composition.

[0014] As a further improvement of the present invention, the following steps are included:

[0015] S1. Wash, dry, and crush soybeans and black beans to obtain bean powder, boil with water, cool to room temperature, add snail enzyme to hydrolyze and inactivate the enzyme to obtain a bean product with resistance substances removed;

[0016] S2. The kudzu vine and Spirulina platensis were washed, dried, crushed, frozen in liquid nitrogen, and returned to room temperature. The bean product obtained in step S2 to remove resistance substances, protein mulberry, was added to water, sterilized, inoculated with selenium-enriched yeast and Aspergillus oryzae seed solution, fermented, filtered, and the filtrate was subjected to ammonium sulfate fractionation precipitation to obtain protein peptides.

[0017] S3. The protein peptide obtained in step S2 was dissolved in water, zinc salt and iron salt were added, the reaction was stirred, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0018] S4. Activating taurine and lysine with N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide, and then reacting the chelated Zn / Fe protein peptide obtained in step S3 to obtain an active protein peptide;

[0019] S5. Pullulan was dissolved in water, the pH of the solution was adjusted to alkaline, maleic anhydride was added, the reaction was heated with stirring, the pH of the solution was adjusted to acidic to terminate the reaction, ethanol precipitation was performed, washing, and drying to obtain carboxylated pullulan;

[0020] S6. The long-chain alkyl acid is reacted with the carboxylated pullulan obtained in step S5 under a catalyst to obtain a hydrophobically modified carboxylated pullulan;

[0021] S7. Add hydrophobically modified carboxylated pullulan and sodium alginate to water, add the active protein peptide, cottonseed protein and lecithin prepared in step S4, stir and mix evenly, add dropwise to the fish oil, emulsify, add dropwise calcium chloride solution, solidify at room temperature, centrifuge, wash, and dry to obtain a plant protein composition.

[0022] As a further improvement of the present invention, the mass ratio of soybean, black bean and snail enzyme in step S1 is 10-15:5-7:1-2, the enzymatic hydrolysis temperature is 40-50°C, the time is 1-3h, and the boiling time of adding water is 20-40min.

[0023] As a further improvement of the present invention, the mass ratio of the kudzu vine, the platen spirulina, the bean product with resistance substances removed, the protein mulberry and the water in step S2 is 10-15:6-8:4-6:1-2:150-200, the inoculation amount of the selenium-enriched yeast and the Aspergillus oryzae seed solution is 1-3v / v% and 2-3v / v%, respectively, and the bacterial content of the seed solution is 10 9 -10 10 cfu / mL, and the fermentation culture conditions are 45-50°C, 100-200r / min, and the fermentation culture is 56-72h.

[0024] As a further improvement of the present invention, the mass ratio of the protein peptide, zinc salt and iron salt in step S3 is 50:3-5:2-4, the stirring reaction time is 30-40 minutes, the zinc salt is selected from at least one of zinc chloride, zinc sulfate and zinc nitrate, and the iron salt is selected from at least one of ferric chloride, ferric sulfate and ferric nitrate.

[0025] As a further improvement of the present invention, the mass ratio of taurine, lysine, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide and chelated Zn / Fe protein peptide in step S4 is 12-13:14-15:21-25:38-42:100-120, the activation temperature is 0-4°C, the time is 30-40min, and the condensation reaction time is 7-10h; the mass ratio of pullulan and maleic anhydride in step S5 is 10:2-3, the pH value of the adjustment solution is alkaline, specifically 8.5-9.5, the pH value of the adjustment solution is acidic, specifically 5.7-6.2, the temperature of the heating and stirring reaction is 35-45°C, and the time is 8-10h.

[0026] As a further improvement of the present invention, the long-chain alkyl acid in step S6 is selected from at least one of octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, and octadecanoic acid, the mass ratio of the long-chain alkyl acid, carboxylated pullulan, and the catalyst is 10-12:100-110:1-2, the catalyst is concentrated sulfuric acid, the reaction temperature is 120-130°C, and the reaction time is 3-5h.

[0027] As a further improvement of the present invention, the mass ratio of the hydrophobically modified carboxylated pullulan, sodium alginate, active protein peptide, cottonseed protein and lecithin in step S7 is 15-20:7-12:14-18:3-5:1-2, and the room temperature curing time is 20-30 min.

[0028] As a further improvement of the present invention, the present invention specifically comprises the following steps:

[0029] S1. 10-15 parts by weight of soybeans and 5-7 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 20-40 min, cooled to room temperature, and 1-2 parts by weight of snail enzyme were added. The enzyme was hydrolyzed at 40-50 ° C for 1-3 h and the enzyme was inactivated to obtain a bean product from which resistance substances were removed.

[0030] S2. 10-15 parts by weight of Gexian rice algae and 6-8 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 4-6 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1-2 parts by weight of protein mulberry were added, 150-200 parts by weight of water, sterilized, and inoculated with a bacterial content of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, with inoculation amounts of 1-3 v / v% and 2-3 v / v%, respectively, at 45-50°C, 100-200 r / min, for fermentation for 56-72 hours, filtering, and subjecting the filtrate to ammonium sulfate graded precipitation to obtain protein peptides;

[0031] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 3-5 parts by weight of a zinc salt and 2-4 parts by weight of an iron salt were added, the reaction was stirred for 30-40min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0032] S4. 12-13 parts by weight of taurine and 14-15 parts by weight of lysine were added to 500 parts by weight of water, 21-25 parts by weight of N-hydroxysuccinimide and 38-42 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 0-4 ° C for 30-40 min, 100-120 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 7-10 h to obtain an active protein peptide;

[0033] S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 8.5-9.5, 0.2-0.3 parts by weight of maleic anhydride was added, heated to 35-45 ° C, stirred for 8-10h, the pH of the solution was adjusted to 5.7-6.2, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0034] S6. 10-12 parts by weight of a long-chain alkyl acid and 100-110 parts by weight of the carboxylated pullulan prepared in step S5 are added to 500 parts by weight of N,N-dimethylformamide, 1-2 parts by weight of concentrated sulfuric acid as a catalyst are added, heated to 120-130 ° C, and stirred for 3-5h to obtain a hydrophobically modified carboxylated pullulan;

[0035] S7. Add 15-20 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 7-12 parts by weight of sodium alginate to 200 parts by weight of water, add 14-18 parts by weight of the active protein peptide prepared in step S4, 3-5 parts by weight of cottonseed protein and 1-2 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify, add dropwise 20-30 parts by weight of 1-2wt% calcium chloride solution, solidify at room temperature for 20-30 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0036] The present invention further protects a plant protein composition obtained by the above production method.

[0037] The present invention has the following beneficial effects:

[0038] Legumes are rich in protein and are excellent plant-based protein raw materials. However, their poor palatability and the widespread presence of anti-nutritional factors such as antigenic proteins, lectins, and non-starch polysaccharides limit their widespread use. Boiling increases the free isoflavone content, boosting the product's isoflavone content and promoting digestion and absorption by animals. Boiling also disrupts the protein's tight quaternary structure, significantly improving digestibility and reducing the activity of anti-nutritional substances. Snail enzymes are a blend of enzymes extracted from the snail's bursa and digestive tract. They contain over 20 enzymes, including cellulase, pectinase, amylase, and protease. These enzymes can enzymatically degrade anti-nutritional factors in legumes, significantly reducing their toxicity and significantly improving the nutritional quality of the product.

[0039] Spirulina gesneri and Spirulina platensis are a type of freshwater algae rich in protein, but they have thick cell walls that are difficult to break, making protein extraction unsafe. The present invention uses liquid nitrogen freezing to form sharp ice knives in the cells, which are convenient for piercing the cell walls, thereby promoting protein extraction in the cells. At the same time, fermentation is carried out using fermentative bacteria, which can degrade and utilize the cell walls as polysaccharides, thereby promoting the disintegration of the cell walls and allowing the full extraction of algal protein. Under the action of selenium-rich yeast Aspergillus oryzae, the obtained protein peptides contain a high content of selenium. Under the action of enzymes produced by the fermentative bacteria, the protein peptides have good activity and are very effective in improving poultry immunity, increasing meat production, and promoting milk production.

[0040] After the protein peptide prepared by the present invention chelates Zn and Fe, the product greatly improves the effect of enhancing animal immunity, has an anti-premature aging effect, has the efficacy of enriching blood, promotes growth and development, improves the food intake of animals, and promotes their rapid growth.

[0041] The chelated Zn / Fe protein peptide prepared by the present invention undergoes a condensation reaction with taurine and lysine to produce an active protein peptide, which enhances the immunity of animals, increases the content of taurine and lysine in animal meat, and improves the nutritional value of the meat. After consumption, it can improve memory, lower blood sugar and blood lipids, promote brain development, and make the meat more delicious and tender, which is popular among consumers.

[0042] Pullulan is a naturally degradable macromolecule with excellent film-forming properties, but it lacks antibacterial properties and has poor mechanical properties. Carboxylation of pullulan significantly improves its mechanical and antibacterial properties. Furthermore, the resulting carboxylated pullulan forms an encapsulating shell with excellent acid resistance, protecting the encapsulated active protein peptides from gastric acid damage, thereby better protecting the activity of the active protein peptides and providing a good regulatory effect. The resulting carboxylated pullulan is further hydrophobically modified to form an amphiphilic polymer polysaccharide, which self-assembles in water to form micelles. This gives the polysaccharide amphiphilicity, thereby forming a thick and tough interfacial layer at the interface of the emulsion, exhibiting good emulsification properties and emulsion stability.

[0043] The plant protein composition prepared by the present invention has good physiological activity. Through the embedding technology, the storage stability, heat resistance, acid resistance and moisture resistance of the protein peptide are improved, the growth and development of poultry and livestock are promoted, the meat quality is improved, and the application prospect is broad. DETAILED DESCRIPTION

[0044] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0045] Pullulan, with a viscosity of 14500-14600 Pa·s, was purchased from Shandong Mimei Biotechnology Co., Ltd.

[0046] Snail enzyme, with a cell wall breaking rate of 98%, was purchased from Shaanxi Zhenghe Pharmaceutical Bioengineering Co., Ltd.

[0047] Selenium-enriched yeast, 10 billion cfu / g, was purchased from Angel Yeast Co., Ltd.; Aspergillus oryzae, 10 billion cfu / g, was purchased from Guangzhou Zhenwei Microbiology Technology Co., Ltd.

[0048] Selenium-enriched yeast and Aspergillus oryzae seed liquid: inoculate the strains into Gao's medium, activate and culture at 52℃, 150r / min for 24h, and obtain a medium with a bacterial count of 10 9 -10 10 cfu / mL of bacterial seed liquid.

[0049] Example 1

[0050] This embodiment provides a method for producing a plant protein composition, which specifically includes the following steps:

[0051] S1. 10 parts by weight of soybeans and 5 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 20 minutes. The mixture was cooled to room temperature. 1 part by weight of snail enzyme was added and enzymatically hydrolyzed at 40°C for 1 hour. The enzyme was inactivated to obtain a bean product from which resistance substances were removed.

[0052] S2. 10 parts by weight of Gexian rice algae and 6 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 4 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1 part by weight of protein mulberry were added, 150 parts by weight of water were sterilized, and the inoculation content was 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 1 v / v% and 2 v / v%, respectively, at 45°C, 100 rpm, fermented for 56 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0053] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 3 parts by weight of zinc chloride and 2 parts by weight of ferric chloride were added, the reaction was stirred for 30 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0054] S4. 12 parts by weight of taurine and 14 parts by weight of lysine were added to 500 parts by weight of water, 21 parts by weight of N-hydroxysuccinimide and 38 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring and activation at 0 ° C for 30 min, 100 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 7-10 h to obtain an active protein peptide;

[0055] S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 8.5, 0.2 parts by weight of maleic anhydride was added, heated to 35 ° C, stirred for 8 hours, the pH of the solution was adjusted to 5.7, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0056] S6 10 parts by weight of octadecanoic acid and 100 parts by weight of the carboxylated pullulan prepared in step S5 were added to 500 parts by weight of N, N- dimethylformamide, 1 part by weight of concentrated sulfuric acid as a catalyst was added, heated to 120 ° C, and the reaction was stirred for 3h to obtain a hydrophobically modified carboxylated pullulan;

[0057] S7. Add 15 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 7 parts by weight of sodium alginate to 200 parts by weight of water, add 14 parts by weight of the active protein peptide prepared in step S4, 3 parts by weight of cottonseed protein and 1 part by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 20 parts by weight of 1wt% calcium chloride solution, solidify at room temperature for 20 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0058] Example 2

[0059] This embodiment provides a method for producing a plant protein composition, which specifically includes the following steps:

[0060] S1. 15 parts by weight of soybeans and 7 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 40 min, cooled to room temperature, and 2 parts by weight of snail enzyme were added. The enzyme was hydrolyzed at 50 ° C for 3 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0061] S2. 15 parts by weight of Gexian rice algae and 8 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 6 parts by weight of the bean product obtained in step S2 to remove resistance substances, 2 parts by weight of protein mulberry were added, 200 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 3 v / v% and 3 v / v%, respectively, at 50°C, 200 r / min, fermented for 72 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0062] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 5 parts by weight of zinc nitrate and 4 parts by weight of ferric nitrate were added, the reaction was stirred for 40 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0063] S4. 13 parts by weight of taurine and 15 parts by weight of lysine were added to 500 parts by weight of water, 25 parts by weight of N-hydroxysuccinimide and 42 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 4 ° C for 40 min, 120 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 10 h to obtain an active protein peptide;

[0064] S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 9.5, 0.3 parts by weight of maleic anhydride was added, heated to 45 ° C, stirred for 10 hours, the pH of the solution was adjusted to 6.2, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0065] S6. 12 parts by weight of myristic acid and 110 parts by weight of the carboxylated pullulan prepared in step S5 were added to 500 parts by weight of N,N-dimethylformamide, 2 parts by weight of concentrated sulfuric acid as a catalyst were added, heated to 130 ° C, and stirred for 5h to obtain a hydrophobically modified carboxylated pullulan;

[0066] S7. Add 20 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 12 parts by weight of sodium alginate to 200 parts by weight of water, add 18 parts by weight of the active protein peptide prepared in step S4, 5 parts by weight of cottonseed protein and 2 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 30 parts by weight of 2wt% calcium chloride solution, solidify at room temperature for 30 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0067] Example 3

[0068] This embodiment provides a method for producing a plant protein composition, which specifically includes the following steps:

[0069] S1. 12 parts by weight of soybeans and 6 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 30 min, cooled to room temperature, and 1.5 parts by weight of snail enzyme was added. The enzyme was hydrolyzed at 45 ° C for 2 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0070] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 2 v / v% and 2.5 v / v%, respectively, at 47°C, 150 r / min, fermented for 64 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0071] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 4 parts by weight of zinc sulfate and 3 parts by weight of ferric sulfate were added, the reaction was stirred for 35 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0072] S4. 12.5 parts by weight of taurine and 14.5 parts by weight of lysine were added to 500 parts by weight of water, 23 parts by weight of N-hydroxysuccinimide and 40 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 2 ° C for 35 min, 110 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 8.5 h to obtain an active protein peptide;

[0073] S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 9, 0.25 parts by weight of maleic anhydride was added, heated to 40 ° C, stirred for 9 hours, the pH of the solution was adjusted to 6, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0074] S6. 11 parts by weight of lauric acid and 105 parts by weight of the carboxylated pullulan prepared in step S5 were added to 500 parts by weight of N,N-dimethylformamide, 1.5 parts by weight of concentrated sulfuric acid as a catalyst were added, heated to 125 ° C, and stirred for 4h to obtain a hydrophobically modified carboxylated pullulan;

[0075] S7. Add 17 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the active protein peptide prepared in step S4, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0076] Comparative Example 1

[0077] Compared with Example 3, the difference is that no helicase is added in step S1.

[0078] The details are as follows:

[0079] S1. Wash 12 parts by weight of soybeans and 6 parts by weight of black beans, dry them, and crush them to obtain bean powder. Add 200 parts by weight of water and boil them for 30 minutes. Cool them to room temperature to obtain a bean product from which the resistance substances have been removed.

[0080] Comparative Example 2

[0081] Compared with Example 3, the difference is that selenium-enriched yeast is not inoculated in step S2.

[0082] The details are as follows:

[0083] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of Aspergillus oryzae seed solution, with an inoculation amount of 4.5 v / v%, 47° C., 150 r / min, fermentation culture for 64 hours, filtration, and the filtrate is subjected to ammonium sulfate graded precipitation to obtain protein peptides.

[0084] Comparative Example 3

[0085] Compared with Example 3, the difference is that Aspergillus oryzae is not inoculated in step S2.

[0086] The details are as follows:

[0087] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast seed liquid, with an inoculation amount of 4.5v / v%, 47°C, 150r / min, fermentation culture for 64h, filtration, and the filtrate is subjected to ammonium sulfate graded precipitation to obtain protein peptides.

[0088] Comparative Example 4

[0089] Compared with embodiment 3, the difference is that step S3 is not performed.

[0090] The details are as follows:

[0091] S1. 12 parts by weight of soybeans and 6 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 30 min, cooled to room temperature, and 1.5 parts by weight of snail enzyme was added. The enzyme was hydrolyzed at 45 ° C for 2 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0092] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 2 v / v% and 2.5 v / v%, respectively, at 47°C, 150 r / min, fermented for 64 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0093] S3. 12.5 parts by weight of taurine and 14.5 parts by weight of lysine were added to 500 parts by weight of water, 23 parts by weight of N-hydroxysuccinimide and 40 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring and activation at 2 ° C for 35 min, 110 parts by weight of the protein peptide prepared in step S2 was added, and the condensation reaction was carried out at room temperature for 8.5 h to obtain an active protein peptide;

[0094] S4. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 9, 0.25 parts by weight of maleic anhydride was added, heated to 40 ° C, stirred for 9 hours, the pH of the solution was adjusted to 6, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0095] S5. 11 parts by weight of lauric acid and 105 parts by weight of the carboxylated pullulan prepared in step S4 were added to 500 parts by weight of N,N-dimethylformamide, 1.5 parts by weight of concentrated sulfuric acid as a catalyst were added, heated to 125 ° C, and the reaction was stirred for 4h to obtain a hydrophobically modified carboxylated pullulan;

[0096] S6. Add 17 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S5 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the active protein peptide prepared in step S3, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0097] Comparative Example 5

[0098] Compared with embodiment 3, the difference is that step S4 is not performed.

[0099] The details are as follows:

[0100] S1. 12 parts by weight of soybeans and 6 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 30 min, cooled to room temperature, and 1.5 parts by weight of snail enzyme was added. The enzyme was hydrolyzed at 45 ° C for 2 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0101] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 2 v / v% and 2.5 v / v%, respectively, at 47°C, 150 r / min, fermented for 64 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0102] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 4 parts by weight of zinc sulfate and 3 parts by weight of ferric sulfate were added, the reaction was stirred for 35 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0103] S4. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 9, 0.25 parts by weight of maleic anhydride was added, heated to 40 ° C, stirred for 9 hours, the pH of the solution was adjusted to 6, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0104] S5. 11 parts by weight of lauric acid and 105 parts by weight of the carboxylated pullulan prepared in step S4 were added to 500 parts by weight of N,N-dimethylformamide, 1.5 parts by weight of concentrated sulfuric acid as a catalyst were added, heated to 125 ° C, and the reaction was stirred for 4h to obtain a hydrophobically modified carboxylated pullulan;

[0105] S6. Add 17 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S5 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0106] Comparative Example 6

[0107] Compared with embodiment 3, the difference is that step S5 is not performed.

[0108] The details are as follows:

[0109] S1. 12 parts by weight of soybeans and 6 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 30 min, cooled to room temperature, and 1.5 parts by weight of snail enzyme was added. The enzyme was hydrolyzed at 45 ° C for 2 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0110] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 2 v / v% and 2.5 v / v%, respectively, at 47°C, 150 r / min, fermented for 64 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0111] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 4 parts by weight of zinc sulfate and 3 parts by weight of ferric sulfate were added, the reaction was stirred for 35 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0112] S4. 12.5 parts by weight of taurine and 14.5 parts by weight of lysine were added to 500 parts by weight of water, 23 parts by weight of N-hydroxysuccinimide and 40 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 2 ° C for 35 min, 110 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 8.5 h to obtain an active protein peptide;

[0113] S5. 11 parts by weight of dodecanoic acid and 105 parts by weight of pullulan were added to 500 parts by weight of N,N- dimethylformamide, 1.5 parts by weight of concentrated sulfuric acid as a catalyst was added, heated to 125 ° C, and the reaction was stirred for 4h to obtain a hydrophobically modified pullulan;

[0114] S6. Add 17 parts by weight of the hydrophobically modified pullulan prepared in step S5 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the active protein peptide prepared in step S4, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0115] Comparative Example 7

[0116] Compared with embodiment 3, the difference is that step S6 is not performed.

[0117] The details are as follows:

[0118] S1. 12 parts by weight of soybeans and 6 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 30 min, cooled to room temperature, and 1.5 parts by weight of snail enzyme was added. The enzyme was hydrolyzed at 45 ° C for 2 h and the enzyme was inactivated to obtain a bean product with resistance substances removed.

[0119] S2. 12 parts by weight of Gexian rice algae and 7 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 5 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1.5 parts by weight of protein mulberry were added to 170 parts by weight of water, sterilized, and inoculated with a bacterial count of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, inoculated at 2 v / v% and 2.5 v / v%, respectively, at 47°C, 150 r / min, fermented for 64 h, filtered, and the filtrate was subjected to ammonium sulfate graded precipitation to obtain protein peptides;

[0120] S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 4 parts by weight of zinc sulfate and 3 parts by weight of ferric sulfate were added, the reaction was stirred for 35 min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide;

[0121] S4. 12.5 parts by weight of taurine and 14.5 parts by weight of lysine were added to 500 parts by weight of water, 23 parts by weight of N-hydroxysuccinimide and 40 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 2 ° C for 35 min, 110 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 8.5 h to obtain an active protein peptide;

[0122] S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 9, 0.25 parts by weight of maleic anhydride was added, heated to 40 ° C, stirred for 9 hours, the pH of the solution was adjusted to 6, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan;

[0123] S6. Add 17 parts by weight of the carboxylated pullulan prepared in step S5 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the active protein peptide prepared in step S4, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0124] Comparative Example 8

[0125] Compared with Example 3, the difference is that no hydrophobically modified carboxylated pullulan is added in step S7.

[0126] The details are as follows:

[0127] S7. Add 27 parts by weight of sodium alginate to 200 parts by weight of water, add 16 parts by weight of the active protein peptide obtained in step S4, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0128] Comparative Example 9

[0129] Compared with Example 3, the difference is that sodium alginate is not added in step S7.

[0130] The details are as follows:

[0131] S7. Add 27 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 to 200 parts by weight of water, add 16 parts by weight of the active protein peptide prepared in step S4, 4 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0132] Comparative Example 10

[0133] Compared with Example 3, the difference is that no active protein peptide is added in step S7.

[0134] The details are as follows:

[0135] S7. Add 17 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 10 parts by weight of sodium alginate to 200 parts by weight of water, add 20 parts by weight of cottonseed protein and 1.5 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify at 10,000 r / min for 15 minutes, add dropwise 25 parts by weight of 1.5wt% calcium chloride solution, solidify at room temperature for 25 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

[0136] Test Example 1

[0137] The plant protein compositions prepared in Examples 1-3 of the present invention and Comparative Examples 1-10 were subjected to performance tests. The results are shown in Table 1.

[0138] The crude protein (CP) and crude fat (EE) contents were determined according to the AOAC (AOAC. Official methods of analysis of AOAC [S]. Arlington: Association of Analytical Chemists, 2005.) method.

[0139] The total amino acid content was determined in accordance with GB / T 18246-2019. The feed samples were treated with performic acid oxidation and hydrolysis method and determined using a Hitachi L-8900 fully automatic amino acid analyzer.

[0140] The selenium content was determined according to GB5009.93-2017 method.

[0141] Table 1

[0142]

[0143] As can be seen from the above table, the plant protein compositions prepared in Examples 1-3 of the present invention are rich in protein and free amino acids, have a low crude fat content, and a high selenium content.

[0144] Test Example 2

[0145] 420 12-week-old young chickens of similar body weight were selected and divided into 14 treatments (a control group, Example 1-3 groups, and Comparative Example 1-10 groups), with 3 replicates for each treatment and 10 chickens per replicate, divided into two cages. The preliminary test lasted 7 days and the final test lasted 28 days. The chickens were raised in fully enclosed cages with three layers of fully overlapping cages. Feeding management and immunization were carried out according to routine procedures. In the fourth week of the experiment, feeding was restricted according to actual body weight. No chickens died during the feeding process. The experimental diet was composed of the following (mass percentage): 65.4% corn, 23.4% soybean meal, 4.84% wheat bran, 0.57% wheat flour, 1.45% calcium hydrogen phosphate, 2.36% stone powder, 0.28% salt, 0.08% methionine, 0.12% choline, 0.1% baking soda, 0.8% zeolite powder, 0.1% chlortetracycline, and 0.5% premix. 0.05 wt% of the corresponding prepared plant protein composition was added to the feed of Example 1-3 and Comparative Example 1-10 groups, respectively, and 0.05 wt% of corn was added to the feed of the control group.

[0146] Before the start of the experiment in week 0 and after the end of week 4, the average weight of each group of birds was measured and the average daily weight gain was calculated. Feed intake (measured weekly, averaged across replicates, and ultimately compared to the average daily feed intake for the entire period) was also measured. The feed-to-weight ratio was also measured. The results are shown in Table 2.

[0147] Table 2

[0148] Group Average daily weight gain (g) Material-to-weight ratio control group 1.92 23.1 Example 1 4.52 12.8 Example 2 4.61 12.9 Example 3 4.67 11.5 Comparative Example 1 4.43 16.5 Comparative Example 2 4.38 15.9 Comparative Example 3 4.41 15.3 Comparative Example 4 4.21 18.9 Comparative Example 5 3.87 21.2 Comparative Example 6 4.12 16.8 Comparative Example 7 4.01 19.0 Comparative Example 8 4.21 18.4 Comparative Example 9 4.09 19.2 Comparative Example 10 3.87 20.5

[0149] As can be seen from the above table, the plant protein compositions prepared in Examples 1-3 of the present invention can significantly promote the growth of young chickens, reduce feed-to-weight ratio, and improve feed utilization.

[0150] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for producing a plant protein composition, characterized in that: The following steps are involved: S1. Removal of resistance factors in legumes; S2. Fermentation of microalgae with legumes and protein mulberry to separate and obtain protein peptides; S3. Metal ion chelation by protein peptides; S4. Condensation of protein peptides that chelate metal ions; S5. Preparation of carboxylated pullulan; S6. Preparation of hydrophobically modified carboxylated pullulan; S7. Preparation of plant protein composition.

2. The production method according to claim 1, characterized in that The following steps are involved: S1. Wash, dry, and crush soybeans and black beans to obtain bean powder, boil with water, cool to room temperature, add snail enzyme to hydrolyze and inactivate the enzyme to obtain a bean product with resistance substances removed; S2. The kudzu vine and Spirulina platensis were washed, dried, crushed, frozen in liquid nitrogen, and returned to room temperature. The bean product obtained in step S2 to remove resistance substances, protein mulberry, was added to water, sterilized, inoculated with selenium-enriched yeast and Aspergillus oryzae seed solution, fermented, filtered, and the filtrate was subjected to ammonium sulfate fractionation precipitation to obtain protein peptides. S3. The protein peptide obtained in step S2 was dissolved in water, zinc salt and iron salt were added, the reaction was stirred, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide; S4. Activating taurine and lysine with N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide, and then reacting the chelated Zn / Fe protein peptide obtained in step S3 to obtain an active protein peptide; S5. Pullulan was dissolved in water, the pH of the solution was adjusted to alkaline, maleic anhydride was added, the reaction was heated with stirring, the pH of the solution was adjusted to acidic to terminate the reaction, ethanol precipitation was performed, washing, and drying to obtain carboxylated pullulan; S6. The long-chain alkyl acid is reacted with the carboxylated pullulan obtained in step S5 under a catalyst to obtain a hydrophobically modified carboxylated pullulan; S7. Add hydrophobically modified carboxylated pullulan and sodium alginate to water, add the active protein peptide, cottonseed protein and lecithin prepared in step S4, stir and mix evenly, add dropwise to the fish oil, emulsify, add dropwise calcium chloride solution, solidify at room temperature, centrifuge, wash, and dry to obtain a plant protein composition.

3. The production method according to claim 2, characterized in that In step S1, the mass ratio of soybean, black bean and snail enzyme is 10-15:5-7:1-2, the enzymatic hydrolysis temperature is 40-50° C., the time is 1-3 hours, and the boiling time of adding water is 20-40 minutes.

4. The production method according to claim 2, characterized in that In step S2, the mass ratio of the radix serratae, the spirulina platensis, the bean product from which resistance substances are removed, the protein mulberry and the water is 10-15:6-8:4-6:1-2:150-200, the inoculation amount of the selenium-enriched yeast and the Aspergillus oryzae seed solution is 1-3 v / v% and 2-3 v / v%, respectively, and the bacterial content of the seed solution is 10 9 -10 10 cfu / mL, and the fermentation culture conditions are 45-50°C, 100-200r / min, and the fermentation culture is 56-72h.

5. The production method according to claim 2, characterized in that The mass ratio of the protein peptide, zinc salt and iron salt in step S3 is 50:3-5:2-4, the stirring reaction time is 30-40 minutes, the zinc salt is selected from at least one of zinc chloride, zinc sulfate and zinc nitrate, and the iron salt is selected from at least one of ferric chloride, ferric sulfate and ferric nitrate.

6. The production method according to claim 2, characterized in that The mass ratio of taurine, lysine, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide and chelated Zn / Fe protein peptide in step S4 is 12-13:14-15:21-25:38-42:100-120, the activation temperature is 0-4°C, the time is 30-40min, and the condensation reaction time is 7-10h; the mass ratio of pullulan and maleic anhydride in step S5 is 10:2-3, the pH value of the adjusted solution is alkaline, specifically 8.5-9.5, the pH value of the adjusted solution is acidic, specifically 5.7-6.2, the temperature of the heated and stirred reaction is 35-45°C, and the time is 8-10h.

7. The production method according to claim 2, characterized in that In step S6, the long-chain alkyl acid is selected from at least one of octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, and octadecanoic acid, the mass ratio of the long-chain alkyl acid, carboxylated pullulan, and the catalyst is 10-12:100-110:1-2, the catalyst is concentrated sulfuric acid, the reaction temperature is 120-130° C., and the reaction time is 3-5 h.

8. The production method according to claim 2, characterized in that In step S7, the mass ratio of the hydrophobically modified carboxylated pullulan, sodium alginate, active protein peptide, cottonseed protein and lecithin is 15-20:7-12:14-18:3-5:1-2, and the room temperature curing time is 20-30 minutes.

9. The production method according to claim 2, characterized in that The specific steps include: S1. 10-15 parts by weight of soybeans and 5-7 parts by weight of black beans were washed, dried, and crushed to obtain soy flour. 200 parts by weight of water were added and boiled for 20-40 min, cooled to room temperature, and 1-2 parts by weight of snail enzyme were added. The enzyme was hydrolyzed at 40-50 ° C for 1-3 h and the enzyme was inactivated to obtain a bean product from which resistance substances were removed. S2. 10-15 parts by weight of Gexian rice algae and 6-8 parts by weight of Spirulina platensis were washed, dried, crushed, frozen with liquid nitrogen, and returned to room temperature. 4-6 parts by weight of the bean product obtained in step S2 to remove resistance substances, 1-2 parts by weight of protein mulberry were added, 150-200 parts by weight of water, sterilized, and inoculated with a bacterial content of 10 9 -10 10 cfu / mL of selenium-enriched yeast and Aspergillus oryzae seed liquid, with inoculation amounts of 1-3 v / v% and 2-3 v / v%, respectively, at 45-50°C, 100-200 r / min, for fermentation for 56-72 hours, filtering, and subjecting the filtrate to ammonium sulfate graded precipitation to obtain protein peptides; S3 50 parts by weight of the protein peptide obtained in step S2 was dissolved in 200 parts by weight of water, 3-5 parts by weight of a zinc salt and 2-4 parts by weight of an iron salt were added, the reaction was stirred for 30-40min, dialyzed, and freeze-dried to obtain a chelated Zn / Fe protein peptide; S4. 12-13 parts by weight of taurine and 14-15 parts by weight of lysine were added to 500 parts by weight of water, 21-25 parts by weight of N-hydroxysuccinimide and 38-42 parts by weight of 1-ethyl-(3-dimethylaminopropyl)carbodiimide were added, and after stirring at 0-4 ° C for 30-40 min, 100-120 parts by weight of the chelated Zn / Fe protein peptide prepared in step S3 was added, and the condensation reaction was carried out at room temperature for 7-10 h to obtain an active protein peptide; S5. 1 part by weight of pullulan was dissolved in 200 parts by weight of water, the pH of the solution was adjusted to 8.5-9.5, 0.2-0.3 parts by weight of maleic anhydride was added, heated to 35-45 ° C, stirred for 8-10h, the pH of the solution was adjusted to 5.7-6.2, the reaction was terminated, ethanol precipitated, washed, and dried to obtain carboxylated pullulan; S6. 10-12 parts by weight of a long-chain alkyl acid and 100-110 parts by weight of the carboxylated pullulan prepared in step S5 are added to 500 parts by weight of N,N-dimethylformamide, 1-2 parts by weight of concentrated sulfuric acid as a catalyst are added, heated to 120-130 ° C, and stirred for 3-5h to obtain a hydrophobically modified carboxylated pullulan; S7. Add 15-20 parts by weight of the hydrophobically modified carboxylated pullulan prepared in step S6 and 7-12 parts by weight of sodium alginate to 200 parts by weight of water, add 14-18 parts by weight of the active protein peptide prepared in step S4, 3-5 parts by weight of cottonseed protein and 1-2 parts by weight of lecithin, stir and mix evenly, add dropwise to 500 parts by weight of fish oil, emulsify, add dropwise 20-30 parts by weight of 1-2wt% calcium chloride solution, solidify at room temperature for 20-30 minutes, centrifuge, wash, and dry to obtain a plant protein composition.

10. A plant protein composition produced by the production method according to any one of claims 1 to 9.

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