Cottonseed peptide prepared based on multienzyme synergistic directional enzymolysis as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of cottonseed peptides, and particularly relates to a cottonseed peptide prepared based on multienzyme synergistic directional enzymolysis and a preparation method and application thereof. The preparation method for preparing the cottonseed peptide based on multi-enzyme synergistic directional enzymolysis comprises the following steps: (1) crushing cottonseeds to obtain cottonseed powder; (2) uniformly mixing cottonseed meal and water to obtain a mixture, adding compound bacteria into the mixture, fermenting, and sterilizing after fermentation to obtain a fermentation product; (3) continuously adding alkaline protease into the fermentation product, carrying out an enzymolysis reaction, carrying out enzyme deactivation after the reaction is finished, continuously adding bromelain, carrying out an enzymolysis reaction, and carrying out enzyme deactivation after the reaction is finished, so as to obtain an enzymolysis product; and (4) centrifuging the enzymolysis product, collecting supernate, and drying to obtain the cottonseed peptide. The cottonseed peptide prepared by the method can be used for preparing piglet feed and improving gastrointestinal health.

Description

Technical Field

[0001] The present invention belongs to the technical field of cottonseed peptides, and particularly relates to cottonseed peptides prepared based on multi-enzyme coordinated directional enzymatic hydrolysis, and a preparation method and application thereof. Background Art

[0002] Early-weaned piglets have a low resistance to the outside world because their digestive systems are not fully developed, their gastrointestinal functions are weak, and their immune systems are not sound. The weaning process affects their digestion and absorption capacity, which may cause allergic reactions, loss of appetite, low feed conversion rate, and growth retardation in piglets, thereby having a negative impact on the economic benefits of the pig farming industry. Currently, in order to alleviate these problems, the industry is trying to use prebiotics, probiotics, and nucleotides to regulate the intestinal health and enhance immunity of piglets. Chinese patent CN106721247B discloses a feed additive that can promote intestinal development in weaned piglets, reduce diarrhea in weaned piglets, and increase survival rate. However, although these methods have helped to a certain extent, the overall effect is not satisfactory, and further exploration of more effective strategies is still needed to promote the healthy growth of piglets and improve breeding efficiency.

[0003] Cottonseed peptide is a bioactive peptide extracted from cotton seeds. Cotton seeds are rich in protein, which can be converted into cottonseed peptides with specific functional properties through specific enzymatic hydrolysis techniques. It has potential applications in nutritional supplementation, functional food development, and medicine. Currently, there are many types of cottonseed peptides on the market, but there are no products specifically targeted at the health of early-weaned piglets. Piglets have an immature digestive system. If cottonseed peptides are not easily digested and absorbed, then even if they contain a rich source of bioactive peptides, they may not effectively improve their intestinal health or immunity, and may even have the opposite effect.

[0004] Therefore, there is an urgent need for cottonseed peptides prepared based on multi-enzyme coordinated directed enzymatic hydrolysis and their preparation methods and applications. Summary of the Invention

[0005] The purpose of the present invention is to provide a cottonseed peptide prepared based on multi-enzyme coordinated directional enzymolysis, and a preparation method and application thereof.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to obtain cottonseed powder; (2) mixing cottonseed powder and water to obtain a mixture, adding composite bacteria to the mixture to ferment, and sterilizing after the fermentation to obtain a fermentation product; (3) adding alkaline protease to the fermentation product to carry out enzymatic hydrolysis reaction, inactivating the enzyme after the reaction is completed, and then adding bromelain to carry out enzymatic hydrolysis reaction, inactivating the enzyme after the reaction is completed to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product is centrifuged, the supernatant is collected, and dried to obtain cottonseed peptide.

[0007] Furthermore, in step (1), the cotton seeds are crushed to a size of less than 80 mesh.

[0008] Furthermore, the composite bacteria in step (2) include Bacillus licheniformis and Bacillus subtilis.

[0009] In the prior art, cottonseed peptides are typically produced by directly enzymatically hydrolyzing cottonseeds, but these peptides are not ideal for improving piglet diarrhea. The present invention, by fermenting cottonseeds with specific bacteria and then enzymatically hydrolyzing them, produces cottonseed peptides that significantly improve piglet diarrhea. This also reduces the free phenol content in the cottonseed peptides.

[0010] Furthermore, the dosage of the Bacillus licheniformis is (3-5)×10 8 CFU / mL mixture; the amount of Bacillus subtilis is (6-9)×10 8 CFU / mL mixture.

[0011] Furthermore, the fermentation conditions in step (2) are: aerobic fermentation at 35-40°C for 20-25 hours, and an aeration rate of 7.0-7.5 L / min.

[0012] Furthermore, the amount of alkaline protease used in step (3) is 100-110 U / mL of the mixture.

[0013] Furthermore, the amount of bromelain used in step (3) is 110-120 U / mL of the mixture.

[0014] The existing technology generally only uses alkaline protease to enzymatically hydrolyze cottonseed, and the prepared cottonseed peptide has an unsatisfactory effect on improving piglet weight. The present invention first ferments cottonseed and then uses alkaline protease and bromelain to enzymatically hydrolyze it, and the prepared cottonseed peptide is more effective in improving piglet weight.

[0015] Furthermore, the drying conditions in step (4) are: spray drying at a temperature of 80-90°C.

[0016] Furthermore, the centrifugal conditions in step (4) are: rotation speed 6000-6500 rpm / min; centrifugation time is 15-20 min.

[0017] The present invention provides cottonseed peptide prepared by the preparation method based on multi-enzyme coordinated directional enzymolysis.

[0018] The present invention also provides the use of the cottonseed peptide prepared based on multi-enzyme coordinated directional enzymolysis in preparing piglet feed.

[0019] Compared with the prior art, the advantages and beneficial effects of the present invention are: 1. The cottonseed peptide prepared by the present invention through fermentation and enzymatic hydrolysis of specific bacterial pulp cottonseed has excellent effect on improving the diarrhea rate of piglets.

[0020] 2. The present invention first ferments cottonseed and then uses alkaline protease and bromelain to perform enzymatic hydrolysis in sequence. The prepared cottonseed peptide is more effective in improving the weight of piglets.

[0021] 3. The present invention can reduce the content of free phenol in cottonseed peptide through the synergistic effect of fermentation and enzymolysis. DETAILED DESCRIPTION

[0022] 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.

[0023] The raw materials used in the following examples of the present invention are all commercially available commodities: Bacillus licheniformis, product number BMZ135423, Ningbo Mingzhou Biotechnology Co., Ltd.

[0024] Bacillus subtilis, product number BMZ339677, Ningbo Mingzhou Biotechnology Co., Ltd.

[0025] Aspergillus niger, product number B71913, Ningbo Mingzhou Biotechnology Co., Ltd.

[0026] Aspergillus oryzae, BMZ135265, Ningbo Mingzhou Biotechnology Co., Ltd.

[0027] Neutral protease, activity 100 u / mg. Shanghai Yuanye Biotechnology Co., Ltd.

[0028] Alkaline protease, activity 200 u / mg. Shanghai Yuanye Biotechnology Co., Ltd.

[0029] Bromelain, activity 1200 u / mg. Shanghai Yuanye Biotechnology Co., Ltd.

[0030] Ficin, activity 100 u / mg. Shanghai Yuanye Biotechnology Co., Ltd.

[0031] Example 1 This embodiment provides a method for preparing cottonseed peptide based on multi-enzyme coordinated directed enzymatic hydrolysis, comprising the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 37°C for 22 hours, ventilation volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Bacillus licheniformis and Bacillus subtilis. The amount of Bacillus licheniformis was 4×10 8 CFU / mL mixture; the amount of Bacillus subtilis is 7×10 8 CFU / mL mixture.

[0032] (3) Alkaline protease was continued to be added to the fermentation product at a dosage of 105 U / mL of the mixture, and sodium hydroxide was used to adjust the pH to 8, and an enzymatic hydrolysis reaction was carried out at 42°C for 32 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 with lactic acid, and bromelain was continued to be added for enzymatic hydrolysis at 42°C for 18 hours. The dosage of bromelain was 115 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0033] Example 2 This embodiment provides a method for preparing cottonseed peptide based on multi-enzyme coordinated directed enzymatic hydrolysis, comprising the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 35°C for 25 hours, ventilation volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Bacillus licheniformis and Bacillus subtilis. The amount of Bacillus licheniformis was 5×10 8 CFU / mL mixture; the amount of Bacillus subtilis is 6×10 8 CFU / mL mixture.

[0034] (3) Alkaline protease was continued to be added to the fermentation product at a dosage of 110 U / mL of the mixture, and the pH was adjusted to 8 using sodium hydroxide, and an enzymatic hydrolysis reaction was carried out at 40°C for 35 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 using lactic acid, and bromelain was continued to be added for enzymatic hydrolysis at 40°C for 20 hours. The dosage of bromelain was 110 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product. (4) The enzymatic hydrolysis product was centrifuged at a speed of 6500 rpm / min and a centrifugal time of 15 min. The supernatant was collected and spray-dried at 90°C to obtain cottonseed peptide.

[0035] Example 3 This embodiment provides a method for preparing cottonseed peptide based on multi-enzyme coordinated directed enzymatic hydrolysis, comprising the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 40°C for 20 hours, ventilation volume of 7.5 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Bacillus licheniformis and Bacillus subtilis. The amount of Bacillus licheniformis was 3×10 8 CFU / mL mixture; the amount of Bacillus subtilis is 9×10 8 CFU / mL mixture.

[0036] (3) Alkaline protease was continued to be added to the fermentation product at a dosage of 100 U / mL of the mixture, and sodium hydroxide was used to adjust the pH to 8, and an enzymatic hydrolysis reaction was carried out at 45°C for 30 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 with lactic acid, and bromelain was continued to be added for enzymatic hydrolysis at 45°C for 15 hours. The dosage of bromelain was 120 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product was centrifuged at a speed of 6000 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 80°C to obtain cottonseed peptide.

[0037] Comparative Example 1 The difference between this comparative example and Example 1 is that no composite bacteria were used for fermentation.

[0038] The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Mix cottonseed powder and water evenly to obtain a mixture. (3) Alkaline protease was added to the mixture at a dosage of 105 U / mL of the mixture, and sodium hydroxide was used to adjust the pH to 8, and an enzymatic hydrolysis reaction was carried out at 42°C for 32 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 with lactic acid. Bromelain was further added to carry out an enzymatic hydrolysis reaction at 42°C for 18 hours. The dosage of bromelain was 115 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product. (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0039] Comparative Example 2 The difference between this comparative example and Example 1 is that only Bacillus subtilis was used for fermentation.

[0040] The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and Bacillus subtilis was added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 37°C for 22 hours, aeration volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The amount of Bacillus subtilis used was 11×10 8 CFU / mL mixture.

[0041] (3) Alkaline protease was continued to be added to the fermentation product at a dosage of 105 U / mL of the mixture, and sodium hydroxide was used to adjust the pH to 8, and an enzymatic hydrolysis reaction was carried out at 42°C for 32 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 with lactic acid, and bromelain was continued to be added for enzymatic hydrolysis at 42°C for 18 hours. The dosage of bromelain was 115 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0042] Comparative Example 3 The difference between this comparative example and Example 1 is that the types of composite bacteria are different.

[0043] The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 37°C for 22 hours, ventilation volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Aspergillus niger and Aspergillus oryzae. The amount of Aspergillus niger was 4×10 8 CFU / mL mixture; the amount of Aspergillus oryzae was 7×10 8 CFU / mL mixture.

[0044] (3) Alkaline protease was continued to be added to the fermentation product at a dosage of 105 U / mL of the mixture, and sodium hydroxide was used to adjust the pH to 8, and an enzymatic hydrolysis reaction was carried out at 42°C for 32 hours. After the reaction, the enzyme was inactivated, and the pH was adjusted to 7 with lactic acid, and bromelain was continued to be added for enzymatic hydrolysis at 42°C for 18 hours. The dosage of bromelain was 115 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0045] Comparative Example 4 The difference between this comparative example and Example 1 is that only alkaline protease is used for enzymolysis.

[0046] The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 37°C for 22 hours, ventilation volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Bacillus licheniformis and Bacillus subtilis. The amount of Bacillus licheniformis was 4×10 8 CFU / mL mixture; the amount of Bacillus subtilis is 7×10 8 CFU / mL mixture.

[0047] (3) Alkaline protease was further added to the fermentation product at a dosage of 105 U / mL of the mixture, and the pH was adjusted to 8 using sodium hydroxide. The enzymatic hydrolysis reaction was carried out at 42°C for 50 h. After the reaction was completed, the enzyme was inactivated to obtain an enzymatic hydrolysis product. (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0048] Comparative Example 5 The difference between this comparative example and Example 1 is that the types of enzymes are different.

[0049] The preparation method of cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis comprises the following steps: (1) Grinding cotton seeds to less than 80 mesh to obtain cotton seed powder; (2) Cottonseed powder and water were mixed evenly to obtain a mixture, and composite bacteria were added to the mixture for fermentation. The fermentation conditions were as follows: aerobic fermentation at 37°C for 22 hours, ventilation volume of 7.0 L / min, and sterilization after the fermentation to obtain a fermentation product. The composite bacteria included Bacillus licheniformis and Bacillus subtilis. The amount of Bacillus licheniformis was 4×10 8 CFU / mL mixture; the amount of Bacillus subtilis is 7×10 8 CFU / mL mixture.

[0050] (3) Neutral protease was continuously added to the fermentation product at a dosage of 105 U / mL of the mixture, and the pH was adjusted to 7 using sodium hydroxide. The enzymatic hydrolysis reaction was carried out at 42°C for 32 h. After the reaction, the enzyme was inactivated. Ficin was continuously added to the fermentation product for an enzymatic hydrolysis reaction at 42°C for 18 h. The dosage of ficin was 115 U / mL of the mixture. After the reaction, the enzyme was inactivated to obtain an enzymatic hydrolysis product. (4) The enzymatic hydrolysis product was centrifuged at a speed of 6200 rpm / min and a centrifugal time of 20 min. The supernatant was collected and spray-dried at 85°C to obtain cottonseed peptide.

[0051] Performance Testing 1. The cottonseed peptides prepared in Examples 1-3 and Comparative Examples 1-5 were used to determine free gossypol. The detection method was based on the national standard GB13086-2020. The results are shown in Table 1.

[0052] Table 1 Determination results of free gossypol in cottonseed peptides (%)

[0053] 2. Cottonseed peptide was added to the basal diet in Table 2 of Chinese patent CN106721247B (the addition amount was 0.3wt% of the basal diet mass), and commercially available cottonseed peptide (Xi'an Longmao Biotechnology Co., Ltd.) was set as a control group.

[0054] Weaned piglets of similar weight, approximately 30 days old, were randomly divided into nine groups based on initial weight. Each group had six replicates, with 10 pigs per replicate. Each treatment was fed a corresponding diet with free access to food and water throughout the experiment. Feeding and management methods were determined based on the specific conditions of the experimental site.

[0055] The piglets' fasting weights were measured at the start and end of the experiment, and diarrhea was observed and recorded throughout the experiment. Diarrhea rate = number of piglets with diarrhea ÷ total number of piglets × 100%.

[0056] Table 2 Feeding results

[0057] As can be seen from Table 1, the free gossypol content in the cottonseed peptides of Examples 1-2 is low. The effect on weight gain of piglets is better, which is similar to the effect of adding probiotics, etc.; the diarrhea rate is reduced, and the effect is better than adding compound feed additives such as probiotics. The cottonseed peptides prepared by the present invention are rich in a variety of essential and non-essential amino acids. Especially for rapidly growing piglets, these amino acids are the key to building muscle tissue and promoting overall growth. The cottonseed peptides prepared by the present invention are easier to digest and absorb, and are particularly suitable for use in young animals or animals whose digestive systems are not yet fully developed. The specific peptide substances contained in the cottonseed peptides of the present invention also have anti-inflammatory properties, which help to reduce intestinal inflammatory reactions, thereby reducing the incidence of diarrhea. Cottonseed peptides can improve the disease resistance of piglets by enhancing the function of the intestinal mucosal barrier. A good intestinal barrier can effectively prevent pathogenic microorganisms from invading the body, reducing the risk of infection. At the same time, it can indirectly promote the growth of beneficial bacteria by providing nutritional support, maintain the balance of intestinal microecology, and further reduce the occurrence of diarrhea. Cottonseed peptides not only provide a high-quality source of protein, but also provide piglets with a certain amount of energy and other micronutrients, helping them better meet their nutritional needs during growth. Cottonseed peptides can improve feed utilization, allowing piglets to more effectively utilize the nutrients they ingest, thereby promoting weight gain.

[0058] In Comparative Example 1, no composite bacteria were used for fermentation, and the diarrhea rate of piglets became higher and the free phenol content increased.

[0059] In Comparative Example 2, only Bacillus subtilis was used for fermentation, and the diarrhea rate of piglets became higher and the free phenol content increased.

[0060] In Comparative Example 3, the types of composite bacteria were different, the diarrhea rate of piglets became higher, and the free phenol content increased.

[0061] In Comparative Example 4, only alkaline protease was used for enzymatic hydrolysis, and the weight gain of the piglets was reduced and the free phenol content was increased.

[0062] In Comparative Example 5, the types of enzymes were different, the weight gain of the piglets was reduced, and the free phenol content was increased.

[0063] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis, characterized in that: The following steps are involved: (1) Grinding cotton seeds to obtain cottonseed powder; (2) mixing cottonseed powder and water to obtain a mixture, adding composite bacteria to the mixture to ferment, and sterilizing after the fermentation to obtain a fermentation product; (3) adding alkaline protease to the fermentation product to carry out enzymatic hydrolysis reaction, inactivating the enzyme after the reaction is completed, and then adding bromelain to carry out enzymatic hydrolysis reaction, inactivating the enzyme after the reaction is completed to obtain an enzymatic hydrolysis product; (4) The enzymatic hydrolysis product is centrifuged, the supernatant is collected, and dried to obtain cottonseed peptide.

2. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymolysis according to claim 1, characterized in that: In the step (1), the cotton seeds are crushed to less than 80 mesh.

3. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymolysis according to claim 1, characterized in that: The composite bacteria in step (2) include Bacillus licheniformis and Bacillus subtilis.

4. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymolysis according to claim 3, characterized in that: The dosage of the Bacillus licheniformis is (3-5)×10 8 CFU / mL mixture; the amount of Bacillus subtilis is (6-9)×10 8 CFU / mL mixture.

5. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis according to claim 1, characterized in that: The fermentation conditions in step (2) are: aerobic fermentation at 35-40°C for 20-25 hours, and an aeration rate of 7.0-7.5 L / min.

6. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymolysis according to claim 1, characterized in that: The amount of alkaline protease used in step (3) is 100-110 U / mL of the mixture.

7. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymatic hydrolysis according to claim 1, characterized in that: The amount of bromelain used in step (3) is 110-120 U / mL of the mixture.

8. The method for preparing cottonseed peptide based on multi-enzyme coordinated directional enzymolysis according to claim 1, characterized in that: The drying conditions in step (4) are: spray drying at a temperature of 80-90°C.

9. A cottonseed peptide prepared by the preparation method according to any one of claims 1 to 8 based on multi-enzyme coordinated directed enzymatic hydrolysis.

10. Use of the cottonseed peptide prepared by the preparation method according to any one of claims 1 to 8 based on multi-enzyme coordinated directed enzymatic hydrolysis in preparing piglet feed.

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