CN-121801796-B - Hydrolyzed protein for improving intestinal barrier and preventing diarrhea and preparation method thereof

CN121801796BCN 121801796 BCN121801796 BCN 121801796BCN-121801796-B

Abstract

The invention discloses hydrolyzed protein for improving intestinal barrier and preventing diarrhea and a preparation method thereof, belonging to the technical field of microorganisms and enzymes. The invention uses trypsin to carry out enzymolysis on whey protein solution, then uses TG enzyme to carry out crosslinking, finally uses wild protease M to carry out secondary hydrolysis, and finally prepares hydrolyzed protein with diarrhea preventing effect, and further, peptide fragments obtained by screening from the hydrolyzed protein also have the function of improving intestinal barrier. The protein peptide prepared by the method can improve the intestinal barrier function of organisms, reduce osmotic pressure change and achieve the effect of diarrhea prevention.

Inventors

ZHAO WEI
LENG JUNCAI
LIN CHENGJIAN

Assignees

江南大学

Dates

Publication Date: 20260508
Application Date: 20260306

Claims (7)

1. A protein peptide for preventing and assisting in treating diarrhea is characterized in that the protein peptide is IVQKPTPEGDLEILLQK with an amino acid sequence shown in SEQ ID NO. 1, LGEYGFQKFKDLGEEHFK with an amino acid sequence shown in SEQ ID NO. 2 or HLVDEPQKHLVDEPQNLIK with an amino acid sequence shown in SEQ ID NO. 3.
2. A method for preparing a hydrolyzed protein comprising the protein peptide of claim 1, comprising the steps of: (1) Dissolving whey protein in deionized water to obtain a protein solution, adjusting the pH to 7-8, adding 4000-5000U/g trypsin with the mass of whey protein, performing enzymolysis at 35-40 ℃ for 3-4 hours, and inactivating enzyme to obtain a trypsin hydrolyzed protein intermediate product; (2) Adjusting the pH of a trypsin hydrolyzed protein intermediate product to 7-8, adding TG enzyme with the mass of 5-10U/g whey protein for crosslinking for 3-4 hours, adjusting the pH to 8-9 again, adding wild protease M with the mass of 500-1000U/g whey protein for enzymolysis, and inactivating enzyme to obtain hydrolyzed protein; wherein the whey protein is purchased from New Zealand constant natural group and has the model number of WPC 80.
3. The hydrolyzed protein produced by the method of claim 2.
4. A pharmaceutical product comprising the protein peptide of claim 1 or the hydrolyzed protein of claim 3.
5. The medicine according to claim 4, wherein the medicine further comprises pharmaceutically acceptable pharmaceutical excipients, wherein the pharmaceutical excipients refer to conventional pharmaceutical carriers in the pharmaceutical field.
6. The pharmaceutical product according to claim 4, wherein the pharmaceutical product comprises an oral dosage form, an injection dosage form, and an inhalation dosage form.
7. Use of a protein peptide according to claim 1 or a hydrolysed protein according to claim 3 for the preparation of a product for the prevention or co-treatment of diarrhea, characterised in that the product comprises a pharmaceutical product.

Description

Hydrolyzed protein for improving intestinal barrier and preventing diarrhea and preparation method thereof Technical Field The invention relates to hydrolyzed protein for improving intestinal barrier and preventing diarrhea and a preparation method thereof, belonging to the technical field of microorganisms and enzymes. Background Diarrhea is a common symptom of the digestive system, and is mainly caused by various factors such as intestinal barrier injury, osmotic pressure imbalance, pathogenic bacteria infection or dyspepsia and malabsorption. For infants, elderly people with digestive dysfunction, postoperative patients and intestinal dysfunction people, it is important to find safe and effective nutrition intervention means. In the prior art, the use of a single protein source such as whey protein for hydrolysis is a common method for preparing hypoallergenic, easily absorbable formulas, such as formulas for special medical uses. For example, the macromolecular milk proteins are decomposed by hydrolysis technology, which can significantly reduce the sensitization of the macromolecular milk proteins and help reduce the incidence of diarrhea in infants. Whey proteins are believed to have potential for modulating intestinal flora due to their high digestibility and the abundance of bioactive components such as glycomacropeptides. There are products on the market that reduce sensitization by hydrolyzing a single milk protein source. However, these techniques have focused on decreasing sensitization, and there is insufficient research on how to actively repair the intestinal barrier function to prevent and assist in treating diarrhea by decreasing osmotic pressure of the intestinal contents while supplementing nutrients. After some products are supplemented with protein source nutrition, osmotic pressure of intestinal contents is easily increased, and diarrhea of fragile intestinal people is caused. Patent CN109619358A discloses a solid functional beverage based on soybean peptide protein powder and a preparation method thereof, but the patent mainly promotes the growth and propagation of beneficial bacteria of a human body by adding prebiotics such as polysaccharide, cellulose and the like so as to achieve the effect of health care of intestinal tracts of the human body. The prebiotics such as lactitol are not easily digested and absorbed in the stomach and intestine, and still increase osmotic pressure of intestinal contents, resulting in diarrhea. Therefore, the method can supplement nutrients, reduce osmotic pressure of intestinal contents, and actively repair intestinal barrier function so as to achieve the important research significance of preventing and assisting in treating diarrhea. Disclosure of Invention In order to solve the problems, the present application provides a hydrolyzed protein for improving intestinal barrier and preventing diarrhea and a preparation method thereof. The application uses trypsin to carry out enzymolysis on whey protein solution, then uses TG enzyme to carry out crosslinking, finally uses wild protease M to carry out secondary hydrolysis, and finally prepares hydrolyzed protein with diarrhea preventing effect, and further, peptide fragments obtained by screening from the hydrolyzed protein also have the function of improving intestinal barrier. The protein peptide prepared by the method can improve the intestinal barrier function of organisms, reduce osmotic pressure change and achieve the effect of diarrhea prevention. The first object of the invention is to provide an engineering bacterium of escherichia coli, which expresses IVQKPTPEGDLEILLQK with an amino acid sequence shown as SEQ ID NO. 1, LGEYGFQKFKDLGEEHFK with an amino acid sequence shown as SEQ ID NO. 2 or HLVDEPQKHLVDEPQNLIK with an amino acid sequence shown as SEQ ID NO. 3. The second object of the present invention is to provide a protein peptide for preventing and assisting in treating diarrhea, wherein the protein peptide is IVQKPTPEGDLEILLQK with an amino acid sequence shown in SEQ ID NO. 1, LGEYGFQKFKDLGEEHFK with an amino acid sequence shown in SEQ ID NO. 2 or HLVDEPQKHLVDEPQNLIK with an amino acid sequence shown in SEQ ID NO. 3. A third object of the present invention is to provide a method for producing a hydrolyzed protein containing the above-mentioned protein peptide, comprising the steps of: (1) Dissolving whey protein in deionized water to obtain a protein solution, adjusting the pH to 7-8, adding 4000-5000U/g trypsin with the mass of whey protein, performing enzymolysis at 35-40 ℃ for 3-4 hours, and inactivating enzyme to obtain a trypsin hydrolyzed protein intermediate product; (2) Adjusting the pH of the trypsin hydrolyzed protein intermediate product to 7-8, adding TG enzyme with the mass of 5-10U/g whey protein for crosslinking for 3-4 hours, adjusting the pH to 8-9 again, adding wild protease M with the mass of 500-1000U/g whey protein for enzymolysis, and inactivating enzyme to obtain hyd