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Preparation method and application of pelodiscus sinensis collagen peptide

A technology of soft-shelled turtle collagen and collagen peptides, applied in the field of natural active substances, to achieve the effects of increasing peptide content and purity, regulating cellular and humoral immune functions, and strengthening in vitro antioxidant properties

Inactive Publication Date: 2015-11-18
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research reports on the preparation of collagen peptides from soft-shelled turtles as raw materials are rare at home and abroad, and so far there have been no reports on the isolation and extraction of collagen peptides with immune-enhancing activity.

Method used

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  • Preparation method and application of pelodiscus sinensis collagen peptide
  • Preparation method and application of pelodiscus sinensis collagen peptide
  • Preparation method and application of pelodiscus sinensis collagen peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation of embodiment 1 soft-shelled turtle collagen peptide

[0035] A preparation method of soft-shelled turtle collagen peptide, comprising the steps of:

[0036] (1) Pretreatment: wash and cut the skirt of Chinese soft-shelled turtle into pieces, and remove foreign proteins and impurities by treating with phosphate buffer solution with a concentration of 1.25%, which is convenient for enzymolysis. The amount of phosphate buffer added is the skirt of Chinese soft-shelled turtle 10 times the mass;

[0037] (2) Enzymolysis: Add 4000 U / g of papain to the collagen solution obtained in step (1), and enzymolyze it for 4 hours at a temperature of 55°C and a pH value of 7.0, and the extraction rate of collagen peptides is 48%. %;

[0038] (3) Enzyme inactivation: heat in a boiling water bath at 55°C for 10 minutes to inactivate the enzyme;

[0039] (4) Centrifugation: Centrifuge at 4000r / min for 15-20min to obtain supernatant. Take the supernatant;

[0040] (5) ...

Embodiment 2

[0043] Example 2 The Ability of Chinese Soft-shelled Turtle Collagen Peptide to Scavenge Hydroxyl Free Radicals

[0044] The method for measuring hydroxyl radical scavenging ability is: add 2.5mL, 9mmol / LFeSO 4 Solution and 2.5mL, 9mmol / L salicylic acid-ethanol solution, then add 2, 4, 6, 8, 10mg / mL different concentrations of collagen peptide before ultrafiltration and 2.5mL of collagen peptide after ultrafiltration, ultrafiltration Collagen peptides before filtration and collagen peptides after ultrafiltration were all dissolved in pH7.4 phosphate buffer, and then 2.5mL, 8.8mmol / LH 2 o 2 . Use vitamin Vc as a positive control, replace the same volume of phosphate buffer solution with 2.5mL distilled water as the control solution, keep it warm at 37°C for 20min, and use distilled water as the reference solution, calculate: clearance rate (%)=[A 0 -A x ]×100% / A 0 , A 0 is the absorbance value of the blank control solution, A xis the absorbance value after adding the hyd...

Embodiment 3

[0046] Example 3 Antioxidant Ability of Chinese Soft-shelled Turtle Collagen Peptide

[0047] Take 2, 4, 6, 8, 10mg / mL different concentrations of collagen peptides before ultrafiltration and collagen peptides after ultrafiltration 1mL, add 1.25 mg / mL of collagen peptides before ultrafiltration and collagen peptides after ultrafiltration mL, 0.2mol / L, pH7.4 phosphate buffer solution; then add 1mL, 1% potassium ferricyanide solution, mix well, put it in a 50°C water bath for 20min, then add 1.25mL, 10% Trichloroacetic acid, mix well, and centrifuge at 3000r / min for 10min. Take supernatant 2.5mL, add 2.5mL distilled water and 0.5mL, 0.1% FeCl3 solution. Mix well and let it stand for 10 minutes, the system will turn from yellow to green. Use the same concentration of Vc as the control test, use plasma water instead of the sample to adjust the zero, and compare the color at a wavelength of 700nm.

[0048] Since antioxidants (reductants) scavenge free radicals by donating electro...

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Abstract

The invention relates to a preparation method of pelodiscus sinensis collagen peptide. The preparation method is characterized by comprising the following steps: (1) preprocessing; (2) enzymatic hydrolysis: adding papain to a collagen solution obtained from the step (1) for enzymatic hydrolysis so as to convert collagen into collagen peptide; (3) enzyme inactivation; (4) centrifugation: centrifuging enzymatic hydrolysate obtained from the step (3) and reserving supernatant liquid; (5) membrane separation: intercepting a collagen peptide component which is less than 10000Da in molecular weight from the supernatant liquid through membrane separation; (6) concentration; and (7) freeze-drying. Meanwhile, the invention also provides application of the collagen peptide in the preparation of drugs, health care products as well as skin care and beauty products having an immune enhancing function. Compared with the prior art, the preparation method disclosed by the invention has the advantages that the pelodiscus sinensis collagen peptide is extracted within 2-4h only by combining a one-time enzymic method and membrane separation technology, so that enzymatic hydrolysis time is effectively reduced compared with general researches which require 5-6; and in addition, the collagen peptide which is less than 10000Da is obtained through separation, so that peptide content and purity of a finished collagen peptide product are significantly improved.

Description

technical field [0001] The invention relates to a preparation method and application of Chinese soft-shelled turtle collagen peptide, belonging to the field of natural active substances. Background technique [0002] Polysaccharides, proteoglycans, and collagen peptides derived from marine sources are very important natural marine food resources, and often have significant life activities. For example, sea cucumber polysaccharides and seaweed polysaccharides have many research reports, confirming that they have significant biological activities. [0003] Chinese soft-shelled turtle (Pelodiscus sinensis), also known as soft-shelled turtle, has important economic and medicinal value, and is a traditional tonic in my country. "Compendium of Materia Medica", "Chinese Pharmacopoeia of Traditional Chinese Medicine" and other historical works discuss that its function has the dual effects of curing diseases and maintaining health and prolonging life. The soft-shelled turtle also co...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/34C07K1/14A61K38/01A61P37/02A61K8/65
Inventor 李彩燕钱国英许国绿顾璐娜宋伟杨桠楠
Owner ZHEJIANG WANLI UNIV
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