CN116762962A - A polypeptide and applications of the polypeptide and compositions containing the polypeptide - Google Patents

Abstract

The invention discloses a polypeptide, applications of the polypeptide and compositions containing the polypeptide. In the present invention, oyster meat is first subjected to a deodorizing treatment, and a deodorizing film is prepared from polyacrylonitrile, tetrabutyl titanate, glacial acetic acid, and 3-azidopropionic acid. The decomposition and inhibitory effects reduce the fishy substances in the oyster meat, so that the polypeptide composition has a low fishy effect; the deodorized oyster meat undergoes enzymatic treatment of the fishy removal film and spray freeze-drying to obtain oysters with high short peptide content. Peptides make the polypeptide composition easily absorbable; oyster peptides are mixed with other nutrients such as ginseng, polygonatum, sea buckthorn, mulberry, yam and black wolfberry to form a polypeptide composition. The polypeptide composition prepared by the invention has the effects of low fishy smell and easy absorption.

Description

A polypeptide and applications of the polypeptide and compositions containing the polypeptide

Technical field

The present invention relates to the technical field of health care products, specifically a polypeptide, its application and a composition containing the polypeptide.

Background technique

With the rapid development of social economy, aquatic products have become an indispensable part of modern healthy diet structure. Among them, oysters, as the world's largest cultivated shellfish, are rich in polyunsaturated acids, vitamins, and trace elements such as zinc, selenium, and iodine. They are known as the "Ocean of Milk" and "The Brown Rice of the Sea." At present, many researchers have used enzymatic hydrolysis technology to deeply process oysters and developed a variety of health products such as peptide powder and peptide tablets, which are widely used in food preservation due to their good antioxidant and antibacterial activities.

However, oyster peptide products have a strong fishy smell, which seriously affects the quality and acceptability of the product, and has become a technical bottleneck in the promotion and application of functional food and pharmaceutical industries. The main deodorizing methods of oyster peptides include physical methods (adsorption, entrapment and masking), chemical methods (Maillard reaction method and antioxidant method), biological methods (yeast and lactic acid bacteria fermentation) and composite methods. Among them, the physical method is simple and easy to implement, but the effect of deodorizing is limited and will lead to the loss of some nutrients; the chemical method has a more obvious effect of deodorizing, but it is easy to cause chemical residues and deterioration of flavor and color; the biological method has a good effect of deodorizing, However, the dosage of microorganisms needs to be strictly controlled, otherwise odor will be produced due to excessive fermentation, which will affect the nutrition and flavor of the product. The compound method has a better effect of removing odor, but the operation is cumbersome and the cost is high. In addition, oyster short peptides with a relative molecular mass ranging from 180 to 1000 are more compact and easier to be absorbed by the human body than amino acids. However, the traditional preparation process of oyster peptides at this stage generally has a low yield of short peptides. Therefore, how to prepare low-odor oyster peptide products efficiently and non-destructively is one of the hot and cutting-edge research topics in the deep processing of shellfish.

Contents of the invention

The purpose of the present invention is to provide a polypeptide, an application of the polypeptide and a composition containing the polypeptide, so as to solve the problems existing in the prior art.

In order to solve the above technical problems, the present invention provides the following technical solutions:

A polypeptide and its application and a composition containing the polypeptide, the polypeptide is oyster peptide; the polypeptide composition contains oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder and black wolfberry powder.

Further, the oyster peptide is prepared by sequentially undergoing deodorization treatment, enzymatic hydrolysis treatment, and spray freeze-drying treatment of oyster meat.

Further, the deodorizing treatment is as follows: mash the oyster meat and mix it with distilled water, add the deodorizing film, soak it for a period of time, take out the deodorizing film, and prepare the deodorizing oyster meat mixture; the deodorizing film It is prepared from polyacrylonitrile, tetrabutyl titanate, glacial acetic acid and 3-azidopropionic acid.

Further, the enzymatic hydrolysis treatment is as follows: mixing the deodorized oyster meat mixture and mixed enzymes, performing low-temperature ultrasonic extraction and low-temperature centrifugation multiple times, collecting the liquid to obtain the enzymatic hydrolysis filtrate; the mixed enzymes include alkaline protease , trypsin.

Further, the spray freeze-drying process is as follows: atomizing the enzymatic hydrolysis filtrate, spraying it with cold air, and freeze-drying for a period of time to obtain oyster peptides.

Further, a polypeptide and its application and a method for preparing a composition containing the polypeptide include the following preparation steps:

(1) Mix oyster meat and distilled water at a mass ratio of 1:2 to 1:3, soak at 4 to 6°C for 50 to 70 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, and add Distilled water with 5 to 9 times the mass of oyster meat, stir at 50 to 70 rpm for 10 to 20 minutes, then add a deodorizing film with 0.2 to 1.0 times the mass of oyster meat, and deodorize at 50 to 70 rpm for 12 to 24 hours under xenon light. Fish it out to get the fishy oyster meat mixture;

(2) Add a mixed enzyme of 0.05 to 0.15 times the mass of the deodorized oyster meat mixture and distilled water 0.5 to 1.5 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. The mixed enzyme is alkaline. The mass ratio of protease and trypsin is 1:2.3~1:2.7. Add hydrochloric acid until the pH of the enzymatic solution is 6~7. After ultrasonic extraction for 30~60 minutes at 0~10℃, frequency 20kHz and intensity 100~1000W , centrifuge for 10 to 30 minutes at 10000 to 20000 rpm and 0 to 10°C, collect the liquid, repeat the above steps 2 to 4 times for the solid, combine the liquids, and obtain the enzymatic hydrolysis filtrate;

(3) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -30~-70°C, and then freeze-dry it for 5-7 hours at a vacuum of 50~90Pa and -30~-10°C to obtain Oyster peptide; wash ginseng, polygonatum, sea buckthorn, mulberry, yam and black wolfberry with purified water 3 to 5 times respectively, dry them in sterility at 40 to 50°C for 1 to 2 days, cut into small pieces, and grind them into powder respectively. Pass through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder and black wolfberry powder;

(4) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 10:1:2:4:3:2:5:0.5:4 ~20:5:6:8:6:6:10:1.0:4 Mix, stir at 50~70rpm for 10~20min, granulate, and dry at 40~50°C for 12h to obtain a composition containing the polypeptide.

Further, the preparation method of the deodorizing membrane described in step (1) is: combining titanium dioxide fiber membrane, 3-azidopropionic acid, cuprous bromide, hexamethyltriethylenetriamine, N,N-dimethyl Formamide is mixed according to the mass ratio of 5:9:1:11:3~7:11:3:15:3, reacted in a nitrogen atmosphere at 50~60°C for 2~4 hours, and frozen in liquid nitrogen for 30~60 seconds before adding n-hexane until the precipitation is complete, filter, wash with deionized water 3 to 5 times, and vacuum dry at -0.08MPa, 50 to 60°C for 12 hours to obtain a deodorizing film.

Further, the titanium dioxide fiber membrane preparation method is: mixing the fiber membrane and acetic acid mixture at a mass ratio of 0.01:70 to 0.02:70, and the mass ratio of acetic acid and deionized water in the acetic acid mixture is 1:15 to 1: 25. Place it in a polytetrafluoroethylene kettle core, seal the stainless steel autoclave, react at 130 to 180°C for 10 to 14 hours, cool to room temperature, and wash with ethanol and deionized water 3 to 5 times in sequence, until the vacuum degree is 0.084 Dry at MPa and 60°C for 6 hours to obtain a titanium dioxide fiber membrane.

Further, the fiber membrane preparation method is: mix polyacrylonitrile and N,N-dimethylformamide in a mass ratio of 1:10 to 2:10, stir at 200 to 300 rpm for 6 to 8 hours, and then add polypropylene. Tetrabutyl titanate/acetic acid mixture with 0.5 to 1.5 times the mass of nitrile. The mass ratio of tetrabutyl titanate, glacial acetic acid, and N,N-dimethylformamide in the tetrabutyl titanate/acetic acid mixture is 0.3 :0.06:3～0.5:0.07:3, stir at 500～800rpm for 2～3h to obtain the spinning liquid; the spinning liquid is electrospun at a voltage of 10kV, a spinning distance of 15cm and a spinning speed of 1.5mL/h. After spinning, dry it for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a fiber membrane.

Further, the composition containing the polypeptide is applied to oyster peptide lozenges. During application, the composition containing the polypeptide and the starch paste are mixed at a mass ratio of 1:0.2 to 1:0.4, and stirred at 50 to 70 rpm for 20 to 30 minutes before being put in. It is pressed in a tablet press to obtain oyster peptide buccal tablets. The mass ratio of starch and distilled water in the starch paste is 8:10 to 15:10.

Compared with the prior art, the beneficial effects achieved by the present invention are:

In the present invention, oyster peptides are obtained from oyster meat through deodorizing treatment, enzymatic hydrolysis treatment, and spray freeze-drying treatment in sequence, and then blended with other substances to prepare a composition containing polypeptides, which has the effects of low fishy smell and easy absorption.

First of all, a deodorizing film is used in the deodorizing treatment. The deodorizing film is made of polyacrylonitrile, tetrabutyl titanate, glacial acetic acid, and 3-azidopropionic acid; polyacrylonitrile, tetrabutyl titanate, glacial acetic acid After electrospinning into a fiber membrane, under the action of hot acetic acid, tetrabutyl titanate condenses on the fiber surface to form uniformly dispersed titanium dioxide particles. The specific surface area and porosity of the fiber membrane enable the fiber membrane to absorb fishy smell substances. At the same time, using The photooxidation of titanium dioxide decomposes the smelly substances into harmless and odorless substances, so that the polypeptide composition has a low smelly effect; the cyano group in the fiber membrane reacts with the azide group in 3-azidopropionic acid to form triazole Structure, the triazole structure can inhibit the interaction between bacteria and nutrients such as protein and lecithin in oysters to produce fishy substances. At the same time, the carboxyl group in 3-azidopropionic acid can react with fishy substances to reduce the fishy substances and increase the polypeptide composition. Low fishy odor effect.

Secondly, the enzymatic hydrolysis treatment of the present invention uses the steady-state cavitation and mechanical effects generated by low-temperature ultrasonic waves to destroy oyster cells, fully release the active substances in the cells, and promote the full contact between the active substances and the enzyme, improve the activity of the enzyme, and make the protein It fully self-degrades into short peptides with low molecular weight and is easy to absorb. It completely retains the original vitamins, trace elements, taurine and other nutrients of oysters, making the nucleic acid-rich oysters absorbed by the human body faster than single amino acids or proteins after ingestion. Easier to be absorbed by the body; during the spray freeze-drying process, the enzymatic hydrolyzate is atomized and sprayed with low-temperature cold air, which accelerates the drying speed of oyster peptides, rapidly cools and pre-freezes them, and prepares low-density and high-specific surface area Powder particles shorten freeze-drying time; in addition, oyster peptide is mixed with other nutrients such as ginseng, polygonatum, sea buckthorn, mulberry, yam and black wolfberry to form a polypeptide composition, which makes the composition have multi-functional health effects.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention.

In order to more clearly illustrate the method provided by the present invention, the following examples are used to explain in detail. The compositions containing polypeptides and the test methods for each indicator of oyster peptide produced in the following examples are as follows:

Low fishy odor effect: Mix the polypeptide compositions and starch paste prepared in the Examples and Comparative Examples at a mass ratio of 1:0.3, stir at 60 rpm for 25 minutes, and then put them into a tablet press for compression to obtain oyster peptide lozenges, which are in the starch paste. The mass ratio of starch to distilled water is 11.5:10. 10 staff members were selected to evaluate the fishy smell of oyster peptide lozenges and scored according to Table 1. The average score is the fishy smell rating;

Table 1

Points Fishy smell score 0 The fishy smell is very strong 1 Strong fishy smell 2 Average fishy smell 3 Little fishy smell 4 Less fishy smell 5 Almost no fishy smell

Easy absorption effect: Take the oyster peptides prepared in the Examples and Comparative Examples of the same quality and test the short peptide and protein content according to GB/T22729, and calculate the short peptide yield according to the following formula:

Short peptide yield (%) = (short peptide content/protein content) × 100%.

Example 1

(1) Mix polyacrylonitrile and N,N-dimethylformamide at a mass ratio of 1:10. After stirring at 200 rpm for 6 hours, add tetrabutyl titanate/acetic acid mixture 0.5 times the mass of polyacrylonitrile. Titanium The mass ratio of tetrabutyl titanate, glacial acetic acid, and N,N-dimethylformamide in the tetrabutyl titanate/acetic acid mixture is 0.3:0.06:3, stir at 500 rpm for 2 hours, and obtain the spinning liquid; spinning liquid After electrospinning at a voltage of 10kV, a spinning distance of 15cm, and a spinning speed of 1.5mL/h, dry it for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a fiber membrane;

(2) Mix the fiber membrane and acetic acid mixture at a mass ratio of 0.01:70. The mass ratio of acetic acid and deionized water in the acetic acid mixture is 1:15. Place it in a polytetrafluoroethylene kettle core and seal the stainless steel autoclave. After reacting for 10 hours at 130°C, cool to room temperature, wash three times with ethanol and deionized water in sequence, and dry for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a titanium dioxide fiber membrane;

(3) Combine titanium dioxide fiber membrane, 3-azidopropionic acid, cuprous bromide, hexamethyltriethylenetriamine, and N,N-dimethylformamide in a mass ratio of 5:9:1:11:3 Mix and react for 2 hours in a nitrogen atmosphere at 50°C. After freezing in liquid nitrogen for 30 seconds, add n-hexane until the precipitation is complete. Filter, wash with deionized water 3 times, and vacuum dry at -0.08MPa and 50°C for 12 hours to obtain the deionized solution. fishy film;

(4) Mix oyster meat and distilled water at a mass ratio of 1:2, soak at 4°C for 50 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, add distilled water 5 times the mass of oyster meat, After stirring at 50 rpm for 10 minutes, add a deodorizing film that is 0.2 times the mass of oyster meat. Under xenon light, deodorize at 50 rpm for 12 hours. Take out the deodorizing film to obtain a mixed solution of deodorized oyster meat;

(5) Add a mixed enzyme of 0.05 times the mass of the deodorized oyster meat mixture and distilled water 0.5 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. The alkaline protease and trypsin in the mixed enzymes The mass ratio is 1:2.3. Add hydrochloric acid until the pH of the enzymatic hydrolyzate is 6. After ultrasonic extraction for 30 minutes at 0°C, a frequency of 20kHz and an intensity of 100W, centrifuge at 10000rpm and 0°C for 10min. Collect the liquid and press the solid Repeat the above steps twice, combine the liquids, and obtain the enzymatic hydrolysis filtrate;

(6) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -30°C, and then freeze-dry it at a vacuum of 50Pa and -30°C for 5 hours to obtain oyster peptide; combine ginseng, polygonatum, sea buckthorn, Mulberry, yam and black wolfberry were washed three times with purified water respectively, dried under sterility and 40°C1, cut into small pieces, and ground into powder respectively and passed through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, and mulberry powder, yam powder and black wolfberry powder;

(7) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 10:5:1:2:4:3:2:5:0.5 :4 Mix, stir at 50 rpm for 10 minutes, granulate, and dry at 40°C for 12 hours to obtain a composition containing the polypeptide.

Example 2

(1) Mix polyacrylonitrile and N,N-dimethylformamide at a mass ratio of 1.5:10. After stirring at 250 rpm for 7 hours, add tetrabutyl titanate/acetic acid mixture that is 1 times the mass of polyacrylonitrile. Titanium The mass ratio of tetrabutyl titanate, glacial acetic acid, and N,N-dimethylformamide in the tetrabutyl titanate/acetic acid mixture is 0.4:0.065:3, stir at 650 rpm for 2.5 hours, and obtain the spinning liquid; After the liquid was electrospun at a voltage of 10kV, a spinning distance of 15cm, and a spinning speed of 1.5mL/h, it was dried at a vacuum of 0.084MPa and 60°C for 6 hours to obtain a fiber membrane;

(2) Mix the fiber membrane and acetic acid mixture at a mass ratio of 0.015:70. The mass ratio of acetic acid and deionized water in the acetic acid mixture is 1:20. Place it in a polytetrafluoroethylene kettle core and seal the stainless steel autoclave. After reacting for 12 hours at 155°C, cool to room temperature, wash 4 times with ethanol and deionized water in sequence, and dry for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a titanium dioxide fiber membrane;

(3) Combine titanium dioxide fiber membrane, 3-azidopropionic acid, cuprous bromide, hexamethyltriethylenetriamine, and N,N-dimethylformamide in a mass ratio of 6:10:2:13:3 Mix and react for 3 hours in a nitrogen atmosphere at 55°C. After freezing in liquid nitrogen for 45 seconds, add n-hexane until the precipitation is complete, filter, wash 4 times with deionized water, and vacuum dry at -0.08MPa and 55°C for 12 hours to obtain the deionized solution. fishy film;

(4) Mix oyster meat and distilled water at a mass ratio of 1:2.5, soak at 5°C for 60 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, add distilled water 7 times the mass of oyster meat, After stirring at 60 rpm for 15 minutes, add a deodorizing film that is 0.6 times the mass of oyster meat. Under xenon light, deodorize at 60 rpm for 18 hours. Take out the deodorizing film to obtain a mixed solution of deodorized oyster meat;

(5) Add a mixed enzyme with 0.1 times the mass of the deodorized oyster meat mixture and distilled water with 1 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. Alkaline protease and trypsin in the mixed enzymes The mass ratio is 1:2.5. Add hydrochloric acid until the pH of the enzymatic solution is 6.5. After ultrasonic extraction for 45 minutes at 5°C, a frequency of 20kHz and an intensity of 550W, centrifuge at 15000rpm and 5°C for 20min. Collect the liquid and press the solid Repeat the above steps three times, combine the liquids, and obtain the enzymatic hydrolysis filtrate;

(6) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -50°C, and then freeze-dry it at a vacuum of 70Pa and -20°C for 6 hours to obtain oyster peptide; combine ginseng, polygonatum, sea buckthorn, Mulberry, yam and black wolfberry were washed 4 times with pure water respectively, dried under sterility and 45°C for 1.5 days, cut into small pieces, and ground into powder respectively and passed through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, Mulberry powder, yam powder and black wolfberry powder;

(7) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 15:3:4:5:4.5:4:7.5:0.75:4 Mix, stir at 60 rpm for 15 minutes, granulate, and dry at 45°C for 12 hours to obtain a composition containing the polypeptide.

Example 3

(1) Mix polyacrylonitrile and N,N-dimethylformamide at a mass ratio of 2:10. After stirring at 300 rpm for 8 hours, add tetrabutyl titanate/acetic acid mixture that is 1.5 times the mass of polyacrylonitrile. Titanium The mass ratio of tetrabutyl titanate, glacial acetic acid, and N,N-dimethylformamide in the tetrabutyl titanate/acetic acid mixture is 0.5:0.07:3. Stir at 800 rpm for 3 hours to obtain a spinning liquid; spinning liquid After electrospinning at a voltage of 10kV, a spinning distance of 15cm, and a spinning speed of 1.5mL/h, dry it for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a fiber membrane;

(2) Mix the fiber membrane and acetic acid mixture at a mass ratio of 0.02:70. The mass ratio of acetic acid and deionized water in the acetic acid mixture is 1:25. Place it in a polytetrafluoroethylene kettle core and seal the stainless steel autoclave. After reacting for 14 hours at 180°C, cool to room temperature, wash 5 times with ethanol and deionized water in sequence, and dry for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a titanium dioxide fiber membrane;

(3) Combine titanium dioxide fiber membrane, 3-azidopropionic acid, cuprous bromide, hexamethyltriethylenetriamine, and N,N-dimethylformamide in a mass ratio of 7:11:3:15:3 Mix and react for 4 hours in a nitrogen atmosphere at 60°C. After freezing in liquid nitrogen for 60 seconds, add n-hexane until the precipitation is complete. Filter, wash with deionized water 5 times, and vacuum dry at -0.08MPa and 60°C for 12 hours to obtain the deionized solution. fishy film;

(4) Mix oyster meat and distilled water at a mass ratio of 1:3, soak at 6°C for 70 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, add distilled water 9 times the mass of oyster meat, After stirring at 70 rpm for 20 minutes, add a deodorizing film that is 1.0 times the mass of oyster meat. Under xenon light, deodorize at 70 rpm for 24 hours. Take out the deodorizing film to obtain a mixed solution of deodorized oyster meat;

(5) Add a mixed enzyme of 0.15 times the mass of the deodorized oyster meat mixture and distilled water 1.5 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. The alkaline protease and trypsin in the mixed enzymes The mass ratio is 1:2.7. Add hydrochloric acid until the pH of the enzymatic solution is 7. After ultrasonic extraction for 60 minutes at 10°C, a frequency of 20kHz and an intensity of 1000W, centrifuge at 20000rpm and 10°C for 30min. Collect the liquid and press the solid Repeat the above steps 4 times, combine the liquids, and obtain the enzymatic hydrolysis filtrate;

(6) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -70°C, and then freeze-dry it at a vacuum of 90Pa and -10°C for 7 hours to obtain oyster peptide; combine ginseng, polygonatum, sea buckthorn, Mulberry, yam and black wolfberry were washed 5 times with purified water respectively, dried under sterility and 50°C for 2 days, cut into small pieces, and ground into powder respectively and passed through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, and mulberry. powder, yam powder and black wolfberry powder;

(7) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 20:5:6:8:6:6:10:1:4 Mix, stir at 70 rpm for 20 minutes, granulate, and dry at 50°C for 12 hours to obtain a composition containing the polypeptide.

Comparative example 1

(1) Mix polyacrylonitrile and N,N-dimethylformamide at a mass ratio of 1.5:10. After stirring at 250rpm for 7 hours, the voltage is 10kV, the spinning distance is 15cm, and the spinning speed is 1.5mL/h. After electrospinning, dry at a vacuum of 0.084MPa and 60°C for 6 hours to obtain a fiber membrane;

(2) Mix fiber membrane, 3-azidopropionic acid, cuprous bromide, hexamethyltriethylenetriamine, and N,N-dimethylformamide in a mass ratio of 6:10:2:13:3 , react in a nitrogen atmosphere at 55°C for 3 hours, freeze in liquid nitrogen for 45 seconds, add n-hexane until the precipitation is complete, filter, wash 4 times with deionized water, and vacuum dry at -0.08MPa, 55°C for 12 hours to remove the smell. membrane;

(4) Mix oyster meat and distilled water at a mass ratio of 1:2.5, soak at 5°C for 60 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, add distilled water 7 times the mass of oyster meat, After stirring at 60 rpm for 15 minutes, add a deodorizing film with 0.6 times the mass of oyster meat. Under xenon light, deodorize at 50 rpm for 18 hours. Take out the deodorizing film to obtain a mixed solution of deodorizing oyster meat;

(5) Add a mixed enzyme with 0.1 times the mass of the deodorized oyster meat mixture and distilled water with 1 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. Alkaline protease and trypsin in the mixed enzymes The mass ratio is 1:2.5. Add hydrochloric acid until the pH of the enzymatic solution is 6.5. After ultrasonic extraction for 45 minutes at 5°C, a frequency of 20kHz and an intensity of 550W, centrifuge at 15000 rpm and 5°C for 20 minutes. Collect the liquid and solids as described above. Repeat the steps three times, combine the liquids, and obtain the enzymatic hydrolysis filtrate;

(6) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -50°C, and then freeze-dry it at a vacuum of 70Pa and -20°C for 6 hours to obtain oyster peptide; combine ginseng, polygonatum, sea buckthorn, Mulberry, yam and black wolfberry were washed 4 times with pure water respectively, dried under sterility and 45°C for 1.5 days, cut into small pieces, and ground into powder respectively and passed through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, Mulberry powder, yam powder and black wolfberry powder;

(7) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 15:3:4:5:4.5:4:7.5:0.75:4 Mix, stir at 60 rpm for 15 minutes, granulate, and dry at 45°C for 12 hours to obtain a composition containing the polypeptide.

Comparative example 2

The difference between Comparative Example 2 and Example 2 is that there is no step (3), and step (4) is changed to: mix oyster meat and distilled water at a mass ratio of 1:2.5, soak at 5°C for 60 minutes, filter, and repeat the two steps. After several times, beat the oyster meat into a paste, add distilled water with 7 times the mass of the oyster meat, stir at 60 rpm for 15 minutes, add a titanium dioxide fiber membrane with 0.6 times the mass of the oyster meat, and deodorize at 60 rpm for 18 hours under xenon light illumination. Remove the membrane and get the fishy oyster meat mixture. The remaining steps are the same as in Example 2.

Comparative example 3

The difference between Comparative Example 3 and Example 2 is that step (5) is different. Step (5) is changed to: add a mixed enzyme with 0.1 times the mass of the deodorized oyster meat mixture to the deodorized oyster meat mixture and mix it with the deodorized oyster meat. Distilled water with 1 times the mass of the solution was used to obtain the enzymatic hydrolyzate. The mass ratio of alkaline protease and trypsin in the mixed enzyme was 1:2.5. Add hydrochloric acid until the pH of the enzymatic hydrolyzate was 6.5. After extraction for 45 minutes at 5°C, at 15000 rpm, 5 Centrifuge for 20 minutes at ℃, collect the liquid, repeat the above steps three times for the solid, combine the liquids, and obtain the enzymatic hydrolysis filtrate. The remaining steps are the same as in Example 2.

Comparative example 4

The difference between Comparative Example 4 and Example 2 is that step (4) is different. Step (4) is changed to: mix oyster meat and distilled water at a mass ratio of 1:2.5, soak at 5°C for 60 minutes, filter, and repeat the soaking for two times. After several times, beat the oyster meat into a paste, add distilled water with 7 times the mass of the oyster meat, stir at 60 rpm for 15 minutes, add a deodorizing film with 0.6 times the mass of the oyster meat, and deodorize at 60 rpm for 18 hours under no light conditions. Remove the fishy film and get the fishy oyster meat mixture. The remaining steps are the same as in Example 2.

Effect example

Table 2 below shows the performance analysis results of compositions containing polypeptides and oyster peptides using Examples 1 to 3 and Comparative Examples 1 to 4 of the present invention.

Table 2

Fishy smell rating Short peptide yield (%) Example 1 5 54.2 Example 2 5 56.8 Example 3 5 55.7 Comparative example 1 3 55.9 Comparative example 2 4 56.1 Comparative example 3 5 29.5 Comparative example 4 3 56.3

From the comparison of the fishy smell scores of the Examples and Comparative Examples in Table 2, it can be found that the use of a deodorizing film in the deodorizing treatment can significantly reduce the fishy smell of the polypeptide composition. Polyacrylonitrile, tetrabutyl titanate, and glacial acetic acid are treated with Titanium dioxide fiber membrane is prepared by electrospinning and hot acid treatment. Titanium dioxide generates active oxygen under light to decompose fishy substances, and synergizes with the adsorption of the fiber membrane to reduce the fishy substances of oyster peptides, so that the polypeptide composition has a low fishy smell effect. 3-azidopropionic acid further reacts with the fiber membrane to form a triazole structure, and at the same time introduces carboxyl groups. The triazole structure can inhibit the production of fishy smell substances. The carboxyl groups can react with fishy smell substances to reduce fishy smell substances, and can further reduce the polypeptide combination. From the comparison of the fishy smell short peptide yield data of the Examples and Comparative Examples in Table 2, it can be found that the use of low-temperature ultrasonic treatment during the enzymatic hydrolysis of oysters can increase the content of short peptides in the polypeptide composition, making the polypeptide composition Easily absorbable.

It is obvious to those skilled in the art that the present invention is not limited to the details of the above-described exemplary embodiments, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention. Therefore, the embodiments should be regarded as illustrative and non-restrictive from any point of view, and the scope of the present invention is defined by the appended claims rather than the above description, and it is therefore intended that all claims falling within the claims All changes within the meaning and scope of equivalent elements are included in the present invention. Any reference signs in a claim shall not be construed as limiting the claim in question.

Claims (10)

1. A polypeptide and the application of the polypeptide and a composition containing the polypeptide, characterized in that the polypeptide is oyster peptide; the polypeptide composition contains oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder and black wolfberry powder. 2. A polypeptide and the application of the polypeptide and a composition containing the polypeptide according to claim 1, characterized in that the oyster peptide is obtained by sequentially undergoing deodorization treatment, enzymatic hydrolysis treatment and spray freeze-drying treatment of oyster meat. . 3. A polypeptide and the application of the polypeptide and a composition containing the polypeptide according to claim 2, characterized in that the deodorizing treatment is: mash the oyster meat and mix it with distilled water, add a deodorizing film, and soak After a period of time, the fish-removing film is taken out to prepare a mixture of oyster meat-removing liquid; the fish-removing film is prepared from polyacrylonitrile, tetrabutyl titanate, glacial acetic acid, and 3-azidopropionic acid. 4. A kind of polypeptide and the application of the polypeptide and the composition containing the polypeptide according to claim 2, characterized in that the enzymatic hydrolysis treatment is: mixing the deodorized oyster meat mixture and the mixed enzyme, and performing low-temperature ultrasonic extraction. , perform low-temperature centrifugation multiple times, collect the liquid, and obtain an enzymatic hydrolysis filtrate; the mixed enzyme includes alkaline protease and trypsin. 5. A polypeptide and the application of the polypeptide and the composition containing the polypeptide according to claim 2, characterized in that the spray freeze-drying process is: atomizing the enzymatic hydrolysis filtrate and spraying it with cold air, Freeze-dry for a period of time to obtain oyster peptides. 6. A method for preparing polypeptides, applications of polypeptides, and compositions containing polypeptides, which are characterized in that they include the following preparation steps: (1) Mix oyster meat and distilled water at a mass ratio of 1:2 to 1:3, soak at 4 to 6°C for 50 to 70 minutes, filter, repeat the soaking twice, beat the oyster meat into a paste, and add Distilled water with 5 to 9 times the mass of oyster meat, stir at 50 to 70 rpm for 10 to 20 minutes, then add a deodorizing film with 0.2 to 1.0 times the mass of oyster meat, and deodorize at 50 to 70 rpm for 12 to 24 hours under xenon light. Fish it out to get the fishy oyster meat mixture; (2) Add a mixed enzyme of 0.05 to 0.15 times the mass of the deodorized oyster meat mixture and distilled water 0.5 to 1.5 times the mass of the deodorized oyster meat mixture to obtain an enzymatic hydrolyzate. The mixed enzyme is alkaline. The mass ratio of protease and trypsin is 1:2.3~1:2.7. Add hydrochloric acid until the pH of the enzymatic solution is 6~7. After ultrasonic extraction for 30~60 minutes at 0~10℃, frequency 20kHz and intensity 100~1000W , centrifuge for 10 to 30 minutes at 10000 to 20000 rpm and 0 to 10°C, collect the liquid, repeat the above steps 2 to 4 times for the solid, combine the liquids, and obtain the enzymatic hydrolysis filtrate; (3) Atomize the enzymatic hydrolysis filtrate through an atomizer, spray it with air at -30~-70°C, and then freeze-dry it for 5-7 hours at a vacuum of 50~90Pa and -30~-10°C to obtain Oyster peptide; wash ginseng, polygonatum, sea buckthorn, mulberry, yam and black wolfberry with purified water 3 to 5 times respectively, dry them in sterility at 40 to 50°C for 1 to 2 days, cut into small pieces, and grind them into powder respectively. Pass through an 80-mesh sieve to obtain ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder and black wolfberry powder; (4) Oyster peptide, ginseng powder, polygonatum powder, sea buckthorn powder, mulberry powder, yam powder, black wolfberry powder, citric acid, steviol glycosides according to the mass ratio of 10:1:2:4:3:2:5:0.5:4 ~20:5:6:8:6:6:10:1.0:4 Mix, stir at 50~70rpm for 10~20min, granulate, and dry at 40~50°C for 12h to obtain a composition containing the polypeptide. 7. A kind of polypeptide and the application of polypeptide and the preparation method of the composition containing polypeptide according to claim 6, characterized in that the preparation method of the deodorizing film described in step (1) is: titanium dioxide fiber membrane, 3 -Azide propionic acid, cuprous bromide, hexamethyltriethylenetriamine, N,N-dimethylformamide in a mass ratio of 5:9:1:11:3～7:11:3:15: 3. Mix and react for 2 to 4 hours in a nitrogen atmosphere at 50 to 60°C. After freezing in liquid nitrogen for 30 to 60 seconds, add n-hexane until the precipitation is complete, filter, wash with deionized water 3 to 5 times, and freeze at -0.08MPa, Vacuum dry at 50-60°C for 12 hours to obtain a fishy film. 8. A kind of polypeptide and the application of polypeptide and the preparation method of the composition containing the polypeptide according to claim 7, characterized in that the preparation method of the titanium dioxide fiber membrane is: mixing the fiber membrane and acetic acid mixture at a mass ratio of 0.01 :70~0.02:70 mixed, the mass ratio of acetic acid and deionized water in the acetic acid mixture is 1:15~1:25, placed in a polytetrafluoroethylene kettle core, sealed in a stainless steel autoclave, and reacted at 130~180°C After 10 to 14 hours, cool to room temperature, wash 3 to 5 times with ethanol and deionized water in sequence, and dry for 6 hours at a vacuum of 0.084MPa and 60°C to obtain a titanium dioxide fiber membrane. 9. A polypeptide and the application of the polypeptide and the preparation method of the composition containing the polypeptide according to claim 8, characterized in that the fiber membrane preparation method is: polyacrylonitrile, N, N-dimethyl Mix formamide at a mass ratio of 1:10 to 2:10. After stirring for 6 to 8 hours at 200 to 300 rpm, add tetrabutyl titanate/acetic acid mixture that is 0.5 to 1.5 times the mass of polyacrylonitrile. Tetrabutyl titanate/ The mass ratio of tetrabutyl titanate, glacial acetic acid, and N,N-dimethylformamide in the acetic acid mixture is 0.3:0.06:3~0.5:0.07:3. Stir at 500~800rpm for 2~3h to obtain spinning. liquid; after the spinning liquid was electrospun at a voltage of 10kV, a spinning distance of 15cm, and a spinning speed of 1.5mL/h, it was dried at a vacuum of 0.084MPa and 60°C for 6 hours to obtain a fiber membrane. 10. A kind of polypeptide and the application of polypeptide and the preparation method of the composition containing the polypeptide according to claim 6, characterized in that the composition containing the polypeptide is applied to oyster peptide lozenges, and when used, the composition containing the polypeptide is used. The composition and starch paste are mixed at a mass ratio of 1:0.2 to 1:0.4, stirred at 50 to 70 rpm for 20 to 30 minutes, and then put into a tablet press for compression to obtain oyster peptide lozenges. The mass ratio of starch to distilled water in the starch paste is: 8:10～15:10.