Antioxidant polypeptide and preparation method thereof

A technology for antioxidant polypeptides and seeds, applied in the field of bioengineering, can solve problems such as cost reduction, inability to achieve, and complex preparation methods

Pending Publication Date: 2020-04-07

南京平港生物技术有限公司

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AI-Extracted Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides an anti-oxidative polypeptide and its preparation method, which solves the problem that the existing preparation method is relatively complicated and cannot realize the concentrated fermentation and enzymolysis treatment of various proteins, saves production costs, and ...

Abstract

The invention discloses an antioxidant polypeptide and a preparation method thereof. The polypeptide is obtained by carrying out production and processing by using animal serum albumin, corn protein,ovalbumin, wheat gliadin, casein, oil seed protein and soybean protein. The oil seed protein is one or a combination of more of peanut seed protein, sesame seed protein, rapeseed seed protein, sunflower seed protein, cottonseed seed protein, safflower seed protein and coconut seed protein. The invention relates to the technical field of bioengineering. According to the antioxidant polypeptide andthe preparation method thereof, several kinds of proteins can be subjected to centralized fermentation and enzymolysis treatment; the production cost is saved and the preparation process is simple; the objective of reducing the cost by simultaneously carrying out microbial fermentation and enzyme hydrolysis processes is well achieved; and the peptide tail end is modified by terminal peptidase generated by microorganisms, so that the bitter taste of the prepared peptide disappears and the natural fragrance is emitted.

Application Domain

Peptide preparation methodsFermentation

Technology Topic

Sunflower seedSAFFLOWER SEED +22

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Examples

Experimental program(3)

Example Embodiment

[0026] Example 1

[0027] S1. Fermentation enzymatic hydrolysis treatment: first select 7 parts of animal serum albumin, 7 parts of zein, 7 parts of ovalbumin, 7 parts of wheat gliadin, 15 parts of casein, 15 parts of oilseed protein and 15 parts of soybean Pour the protein into the fermenter separately, then add 35mL of warm water, then inoculate the fermentation strains, start the heating equipment, control the temperature of the fermenter at 30℃, and adjust the pH in the range of 6, and then add 16mL of hydrolysis to the fermenter The protease is stirred for 25 minutes at a speed of 250r/min through the stirring device. After the mixing is completed, the fermentation tank is sealed and fermented for 14 hours. The oil seed protein is peanut seed protein, sesame seed protein, rapeseed seed protein, sunflower seed protein, cottonseed seed protein, and red. A composition of flower seed protein and coconut seed protein, and animal serum albumin is a composition of porcine serum albumin, goat serum albumin and bovine serum albumin;

[0028] S2. Filtration treatment: transfer the materials after fermentation in step S1 to the filtration equipment, filter them through the ultrafiltration membrane, filter the fermentation impurities, and collect the filtered fermentation broth;

[0029] S3. Sterilization treatment: the fermentation broth obtained in step S2 is first subjected to a high-temperature sterilization treatment at 120°C for 13 minutes through a steam sterilization device;

[0030] S4. Purification and identification processing: Then the sterilized fermentation broth is purified by SP-Sephadex C-25 ion exchange chromatography, Sephadex G-50 molecular sieve and RP-HPLC reversed-phase high performance liquid chromatography to obtain a mixed antibody Oxidize the polypeptide, and finally identify the amino acid sequence of each polypeptide with a protein solid-phase sequence analyzer. After the identification is successful, the preparation of the mixed antioxidant polypeptide can be completed.

Example Embodiment

[0031] Example 2

[0032] S1. Fermentation and enzymatic hydrolysis: firstly select 5 parts of animal serum albumin, 5 parts of corn protein, 5 parts of ovalbumin, 5 parts of wheat gliadin, 20 parts of casein, 20 parts of oilseed protein and 20 parts of soybean Pour the protein into the fermenter, then add 30mL of warm water, then inoculate the fermentation strains, start the heating equipment, control the temperature of the fermenter at 26℃, and adjust the pH in the range of 5, then add 12mL of hydrolysis to the fermenter The protease is stirred for 20 minutes at a speed of 200r/min through a stirring device. After the mixing is completed, the fermentor is sealed and fermented for 12 hours. The oil seed protein is peanut seed protein, and the animal serum albumin is porcine serum albumin;

[0033] S2. Filtration treatment: transfer the materials after fermentation in step S1 to the filtration equipment, filter them through the ultrafiltration membrane, filter the fermentation impurities, and collect the filtered fermentation broth;

[0034] S3. Sterilization treatment: the fermentation broth obtained in step S2 is subjected to a high-temperature sterilization treatment at 110°C for 10 minutes through a steam sterilization device;

[0035] S4. Purification and identification processing: The sterilized fermentation broth is then purified by SP-Sephadex C-25 ion exchange chromatography, Sephadex G-50 molecular sieve and RP-HPLC reversed-phase high performance liquid chromatography to obtain a mixed antibody Oxidize the polypeptide, and finally use a protein solid-phase sequence analyzer to identify the amino acid sequence of each polypeptide. After the identification is successful, the preparation of the mixed antioxidant polypeptide can be completed.

Example Embodiment

[0036] Example 3

[0037] S1. Fermentation enzymatic hydrolysis treatment: firstly select 10 parts of animal serum albumin, 10 parts of zein, 10 parts of ovalbumin, 10 parts of wheat gliadin, 10 parts of casein, 10 parts of oilseed protein and 10 parts of soybean Pour the protein into the fermenter separately, then add 40mL of warm water, then inoculate the fermentation bacteria, start the heating equipment, control the temperature of the fermenter at 33℃, and adjust the pH in the range of 7, and then add 20mL of hydrolysis to the fermenter The protease is stirred for 30 minutes at a rotational speed of 300r/min through a stirring device. After the mixing is completed, the fermentation tank is sealed and fermented for 16 hours. The oil seed protein is coconut seed protein, and the animal serum albumin is bovine serum albumin;

[0038] S2. Filtration treatment: transfer the materials after fermentation in step S1 to the filtration equipment, filter them through the ultrafiltration membrane, filter the fermentation impurities, and collect the filtered fermentation broth;

[0039] S3. Sterilization treatment: the fermentation broth obtained in step S2 is first subjected to a high-temperature sterilization treatment at 130°C for 15 minutes through a steam sterilization device;

[0040] S4. Purification and identification processing: The sterilized fermentation broth is then purified by SP-Sephadex C-25 ion exchange chromatography, Sephadex G-50 molecular sieve and RP-HPLC reversed-phase high performance liquid chromatography to obtain a mixed antibody Oxidize the polypeptide, and finally use a protein solid-phase sequence analyzer to identify the amino acid sequence of each polypeptide. After the identification is successful, the preparation of the mixed antioxidant polypeptide can be completed.

[0041] In summary

[0042] The present invention can realize concentrated fermentation and enzymatic hydrolysis treatment of various proteins, save production cost, simple preparation process, and well achieve the purpose of reducing cost by using microbial fermentation and enzymatic hydrolysis processes at the same time. The produced terminal peptidase modifies the end of the peptide, so that the bitter taste of the prepared peptide disappears, and the natural aromatic taste is emitted. The preparation conditions are mild and easy to control, thereby greatly facilitating the preparation of the polypeptide by the producer.

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