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Fish skin derived matrix metalloproteinase antagonist peptide

A matrix metal and protease technology, applied in fermentation and other directions, can solve the problems of high cost, ineffective effect, and low utilization rate of polypeptides, and achieve the effects of reducing environmental pollution, slowing down skin oxidative damage, and important social benefits

Active Publication Date: 2015-11-25
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported at home and abroad that many natural animal and plant extracts have the effect of anti-oxidation and delaying skin photoaging, but their content is small, the utilization rate is low, and the effect is not obvious
Polypeptides with metalloprotease-inhibiting activity synthesized by chemical methods are expensive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Use 300 grams of salmon skin as raw material after thawing, soak in 0.1mol / L NaOH solution (1:6w / v) for 30 minutes at room temperature, and then wash with water until it is neutral. Use 0.02mol / L of H 2 SO 4 Soak the solution (1:6w / v) at room temperature for about 30 minutes, and wash with water to pH 5.5. Add distilled water according to the material-to-liquid ratio 1:2-1:6 (w / v), shake and extract in a 60°C water bath for 4 hours, centrifuge to obtain the supernatant and salmon skin gelatin.

[0021] Adjust the concentration of salmon skin gelatin solution to 20g / L, pH to 7.3, and use a combination of trypsin and alkaline protease (protease produced by Bacillus licheniformis 2709) for hydrolysis, the concentration of trypsin is 1200U / g·prot, alkali The concentration of protease is 1000U / g·prot, and the enzymolysis is performed at 50℃. The enzymolysis process is controlled by an enzyme sensor and an artificial neural network. After 120min of enzymolysis, a quantitative pum...

Embodiment 2

[0025] Take 200g fresh cod skin as raw material, soak in 0.05mol / L NaOH solution (material-to-liquid ratio 1:6w / v) for 40 minutes at room temperature, wash with water to neutral, 0.03mol / L H 2 SO 4 The solution (1:6w / v) was soaked at room temperature for about 30 minutes, and washed with water to pH 6.0. Add distilled water according to the material-to-liquid ratio of 1:4 (w / v), shake and extract in a 65°C water bath for 5 hours, and centrifuge to obtain the supernatant to obtain cod skin gelatin.

[0026] Adjust the concentration of cod skin gelatin solution to 20g / L, adjust the initial pH to 7.5, and use a combination of trypsin and alkaline protease (protease produced by Bacillus licheniformis 2709) for hydrolysis. The concentration of trypsin is 1200U / g·prot. The concentration of protease is 1000U / g·prot, and the enzymolysis is performed at 50℃. The enzymolysis process is controlled by an enzyme sensor and an artificial neural network. After 120min of enzymolysis, a quantitati...

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PUM

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Abstract

The invention provides a preparation method of a polypeptide for inhibiting activity of matrix metalloproteinase in photo-aged skin; by taking fish skin as a raw material, an enzyme sensor-artificial neural network online monitoring model is established through a combined enzyme of alkaline protease and pancreatin to control a hydrolysis progress, so that hydrolysis degree is 18%-19%; an enzymatic hydrolysis solution is filtered by virtue of a microporous filtering membrane of 0.22 micron and active peptide with molecular weight cutoff ranging from 300Da to 1000Da is ultra-filtered by virtue of a spiral-wound ultrafiltration membrane; and finally, the peptide is subject to sequential separation and purification by virtue of an SP-Sephadex C25 cation exchange column and a Sephadex G25 gel column, and the peptide is freeze-dried so as to obtain a target antagonist peptide that a main feature segment is Gly-Pro matrix metalloproteinase (MMPs). The fish skin gelatin derived bioactive peptide prepared by the invention is capable of significantly inhibiting the increase in skin MMPs content caused by ultraviolet ray and is relatively good in anti-oxidative activity; and the antagonist peptide provides a wider prospect for the development and utilization of active peptide which is prepared from fish skin gelatin and has photo-aging resisting activity for skin.

Description

Technical field [0001] The invention belongs to the technical field of functional peptide preparation, and specifically relates to a fish skin-derived matrix metalloproteinase antagonist peptide. Background technique [0002] In recent decades, due to the destruction of the ozone layer and excessive sunbathing, the number of people with photoaging skin has increased. Skin relaxation, roughness, atrophy, wrinkles, pigmentation, telangiectasia, and purpura caused by ultraviolet rays have seriously affected the health of people's skin. Preventing and delaying skin photoaging has become a hot spot in medical research. [0003] Skin photoaging is closely related to the reduction of collagen content in the dermis. Ultraviolet radiation produces too much ROS, which disrupts the balance of the redox system in the body, induces an increase in the level of matrix metalloproteinases (MatrixMetalloProteinasesMMPs), and leads to the degradation of collagen in the dermis. [0004] At present, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
Inventor 侯虎陈铁军樊燕李八方陆姣含王石开司磊磊
Owner OCEAN UNIV OF CHINA
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