Antioxidative peptide and preparation method for same

A technology for separation and purification of antioxidant peptides, applied in the field of food biology, can solve problems such as potential safety hazards and waste of protein resources, and achieve the effect of small molecular weight and strong activity

Inactive Publication Date: 2013-09-04
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Chrysalis is a frog belonging to the genus Maras. Its meat is rich in protein. However, because it may contain a variety of germs and toxins in its body, such as parasites that are extremely harmful to humans have been found in frogs, direct con...

Method used

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  • Antioxidative peptide and preparation method for same
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  • Antioxidative peptide and preparation method for same

Examples

Experimental program
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Effect test

Embodiment 1

[0032] (1) Preparation of cyanotrophin enzymatic hydrolyzate: mix qingyang meat and water at a mass ratio of 1:1, add 0.5% Papain based on the mass of cyanotrophin, enzymatically hydrolyze at 55 °C for 4 h; sterilize at 95 °C Enzyme for 15 min, centrifuge, and take the supernatant to obtain the enzymatic hydrolyzate of cyanotrophin, the degree of hydrolysis is 16.07±0.21%;

[0033] (2) Separation and purification of antioxidant peptides: pass the Qingyang protein enzymatic hydrolyzate through ultrafiltration membranes with a molecular weight cut-off of 50,000 Da and 1,000 Da, filter out macromolecules, take the permeate, and freeze-dry it into powder; exchange it with weak acid cations The resin Amberlite IRC50 was eluted with 50mM ammonium acetate buffer solution with pH values ​​of 5, 10, and 12 respectively, the flow rate was 4mL / min, and the detection wavelength was 214nm. A total of 3 peaks were collected, and the antioxidant activity of each elution peak was determined. ...

Embodiment 2

[0036] (1) Preparation of cyanotrophin enzymatic hydrolyzate: mix qingyang meat with water at a mass ratio of 1:4, add 0.5% flavor enzyme based on the mass of cyanotrophin, enzymolyze at 50°C for 20 hours; sterilize at 90°C Enzyme for 30 minutes, centrifuge, take the supernatant to obtain the enzymatic hydrolyzate of Qingyang protein, the degree of hydrolysis is 14.21±0.54%;

[0037] (2) Separation and purification of antioxidant peptides: pass the Qingyang protein enzymatic hydrolyzate through ultrafiltration membranes with a molecular weight cut-off of 5000Da and 1000Da, filter out macromolecules, take the permeate, freeze-dry into powder; use weak acidic cation exchange resin Amberlite IRC50 was eluted with 50mM ammonium acetate buffer solution with pH values ​​of 5, 10, and 12 respectively, the flow rate was 6mL / min, and the detection wavelength was 214nm. Three peaks were collected, and the antioxidant activity of each elution peak was measured. The fraction with the high...

Embodiment 3

[0040] Carry out in the same manner as in Embodiment 2, except that papain and flavor enzymes are used to compound to obtain the enzymatic hydrolyzate of cyanotrophin, and the degree of hydrolysis is increased to 18.93±0.12%.

[0041] Figure 1 is the spectrum of antioxidant peptides separated and purified by Amberlite IRC50 ion exchange chromatography. It can be seen from Figure 1 that peak B has the highest antioxidant activity; Figure 2 is the spectrum of antioxidant peptides separated and purified by Sephadex G-15 gel filtration chromatography. 2 It can be seen that the activity of component B2 is the highest, and that of B3 is slightly worse. Figure 3 is a semi-preparative high performance liquid chromatography separation and purification antioxidant peptide map, it can be seen from Figure 3 that components B2-c and B3-a have the highest activity. Fig. 4 is the map of the antioxidant peptides separated and purified by analytical high performance liquid chromatography, iden...

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Abstract

The invention relates to an antioxidative peptide and a preparation method for the same. A novel antioxidative peptide product is prepared by hydrolyzing hylarana guentheri proteins via an enzymic method to obtain hylarana guentheri protein enzymatic hydrolysate, and performing separation and purification on the hylarana guentheri protein enzymatic hydrolysate by virtue of an ultrafiltration membrane, an ion exchange chromatography, a gel filtration chromatography and a reversed-phase high-performance liquid chromatography, and the amino acid sequences of the product are determined. The amino acid sequences of the product are Leu-Lys(259.1607Da) and Phe-Lys(293.1446Da), as well as have an antioxidative effect, and high oxygen radical absorption capacities which are 7.83 times and 15.78 times than crude peptides; and moreover, the peptide has the characteristics of being low in molecular weight, and capable of being directly absorbed by a human body, thus being capable of being widely applied to the fields of foods, healthcare products, cosmetics, and the like.

Description

technical field [0001] The invention relates to the preparation method and application of two novel antioxidant peptides, belonging to the field of food biotechnology. Background technique [0002] Since British Harman proposed the theory of free radicals in 1956, people have gradually realized that many chronic diseases of the human body, such as decreased immunity, cardiovascular diseases, neurodegenerative disorders, cancer and aging, are related to free radicals produced by oxidation in the body, so antioxidants research has attracted more and more attention. Traditionally used antioxidants are mostly chemically synthesized antioxidants, such as di-tert-butylhydroxytoluene (BHT) and tert-butylhydroxyanisole (BHA), which are widely used because they are cheap and have strong antioxidant capacity, but Its use is restricted due to its potential harm to human health. Therefore, finding and developing safe, non-toxic and highly effective natural antioxidants has become a re...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/16C07K5/06C07K5/065
Inventor 任娇艳顾敏赵谋明崔春
Owner SOUTH CHINA UNIV OF TECH
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