Collagen bioactive peptide, preparation and use thereof
A collagen peptide and collagen technology, which is applied in food preparation, application, animal feed, etc., can solve the problems of not reporting the biological activity of collagen peptides in detail, not seeing the physical and chemical characteristics of collagen peptides, and increasing the number of antibody-producing cells, etc. Achieve the effect of inhibiting the activity of polyphenol oxidase in vitro, high yield and high food safety
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Example Embodiment
[0028] Example 1
[0029] (1) The washed fish skin is mashed, and then treated with 5-15 times of purified water and 5% NaCl solution to remove water-soluble and salt-soluble impurities respectively.
[0030] (2) Treat the fish skin from which impurities have been removed with 20 times 1% pepsin solution (pH2) for 72 hours, centrifuge (5000×g), take the supernatant, add solid NaCl to make the concentration reach 2.5M, centrifuge (5000×g) ×g), take the precipitate to obtain crude collagen.
[0031] (3) Dissolve the crude collagen with 0.5M acetic acid, and use 0.5M acetic acid as the external dialysis fluid for dialysis for 48-72h, until the dialysate is free of Cl - Until detection, undenatured type I collagen has been obtained.
[0032] (4) Disperse collagen in 20 times the amount of pure water, adjust the pH to 8, heat to 50°C, keep for 40 minutes, and then add trypsin (enzyme and substrate concentration of 60 U / g) in proportion to enzymatic hydrolysis 20mins or until the degree...
Example Embodiment
[0035] Example 2
[0036]Disperse the undenatured type I collagen obtained in steps (1), (2), and (3) in Example 1 in 10 times the amount of pure water, adjust the pH to 2, and heat to 60°C, according to the example Add pepsin in the ratio of 1 for enzymatic hydrolysis. The enzymatic hydrolysis time is about 40mins or until the degree of hydrolysis reaches 15%; put the collagen hydrolysate into a dialysis bag with a molecular weight cut-off of 2kD, and conduct dialysis overnight with distilled water as the external fluid. After freeze-drying to obtain collagen active peptide, after separation and purification by Sephadex-G50 chromatography, the molecular weight of the obtained effective component is 5021D.
Example Embodiment
[0037] Example 3
[0038] Disperse the undenatured type I collagen obtained in steps (1), (2), (3) in Example 1 in 10 times the amount of pure water, adjust the pH to 2, and add in the proportions in Example 1. Pepsin is used for enzymolysis, the temperature is controlled at 30℃, and the enzymolysis time is about 40mins or until the degree of hydrolysis reaches 5%; put the collagen hydrolysate into a dialysis bag with a molecular weight cut-off of 2kD, and conduct dialysis overnight with distilled water as the outer fluid , And then freeze-dried to obtain the collagen active peptide, and after separation and purification by Sephadex-G50 chromatography, the molecular weight of the obtained effective component is 5021D.
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