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Cod skin oligopeptides and separation and purification method thereof and application thereof in preparing alpha-glucosidase inhibitor and medicine for resisting type II diabetes

A technology for separation and purification of skin oligopeptides, applied in the field of preparation of α-glucosidase inhibitors and anti-type 2 diabetes drugs, which can solve the problems of undisclosed hypoglycemic mechanism and achieve the effect of small dispersion

Active Publication Date: 2019-05-21
ZHEJIANG ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The currently disclosed fish skin oligopeptides with hypoglycemic effects all follow the mechanism (1) to play a role in regulating blood sugar, and no other hypoglycemic mechanism has been disclosed

Method used

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  • Cod skin oligopeptides and separation and purification method thereof and application thereof in preparing alpha-glucosidase inhibitor and medicine for resisting type II diabetes
  • Cod skin oligopeptides and separation and purification method thereof and application thereof in preparing alpha-glucosidase inhibitor and medicine for resisting type II diabetes
  • Cod skin oligopeptides and separation and purification method thereof and application thereof in preparing alpha-glucosidase inhibitor and medicine for resisting type II diabetes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] (1) Alaska pollock skin 100g, trypsin 0.2g, water 600mL, put in a 1000mL beaker and put it in a 55°C constant temperature water bath, adjust the pH value to 6.0, and start the stirrer to stir the reaction solution, control the stirrer speed to 300rpm , reacted for 8 hours, raised the temperature to 90° C. to inactivate the enzyme for 20 minutes, took out the reaction solution, and filtered it to obtain the filtrate, which was cod skin collagen peptide mixed peptide. The activity test found that the inhibitory activity IC50 of the cod skin collagen peptide mixture peptide to α-glucosidase was 50.4 mg / mL.

[0064] (2) The filtrate is ultrafiltered with a hollow fiber polysulfone ultrafiltration membrane with a molecular weight cut-off of 3000Da, the filtrate is taken, concentrated, and spray-dried to obtain a finished product of cod skin collagen peptide with a molecular weight of figure 1 ).

[0065] Through the activity test, it was found that the inhibitory activity IC...

Embodiment 2

[0095] (1) Alaska pollock skin 200g, trypsin 0.4g, water 1200mL, put in a 2000mL beaker and put it in a constant temperature water bath at 55°C, adjust the pH value to 6.0, and start the stirrer to stir the reaction solution, control the stirrer speed to 300rpm , reacted for 8 hours, raised the temperature to 90°C to inactivate the enzyme for 20 minutes, took out the reaction solution, and filtered it. The obtained cod skin collagen peptide mixed peptide had an IC 50 It was 52.7 mg / mL.

[0096] (2) The filtrate is ultrafiltered with a hollow fiber polysulfone ultrafiltration membrane with a molecular weight cut-off of 3000Da, the filtrate is taken, concentrated, and spray-dried to obtain a finished cod skin collagen peptide with a molecular weight of Figure 7 ).

[0097] (3) Use Re-HPLC technology to separate and separate the components by gel chromatography. The separation conditions are: chromatographic column: Angilent Eclipse XDB-C18 column (250×4.6mm, 5μm) mobile phase: ...

Embodiment 3

[0099] (1) Alaska pollock skin 1000g, trypsin 2g, water 6000mL, put in a self-made stainless steel container and put it in a 55°C constant temperature water bath, adjust the pH value to 6.0, and start the stirrer to stir the reaction solution, control the speed of the stirrer 300rpm, react for 8h, heat up to 90°C to inactivate the enzyme for 20min, take out the reaction solution, filter, and the obtained cod skin collagen peptide mixed peptide has an IC of α-glucosidase inhibitory activity 50 It was 51.9 mg / mL.

[0100] (2) The filtrate is ultrafiltered with a hollow fiber polysulfone ultrafiltration membrane with a molecular weight cut-off of 3000Da, the filtrate is taken, concentrated, and spray-dried to obtain a finished cod skin collagen peptide with a molecular weight of <3000Da. The filtrate is spray-dried to obtain the finished cod skin collagen peptide. Weigh 2g of cod skin collagen peptide, add 2mL of water to dissolve, take 1mL of collagen peptide aqueous solution a...

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Abstract

The invention discloses cod skin oligopeptides and a separation and purification method thereof and an application thereof. The amino acid sequence of the cod skin oligopeptides is Glu-Gly-Gly-Tyr-Thr-Arg, Tyr-Val-Arg or Phe-Tyr-Glu respectively. The separation and purification method comprises the following steps of: preparing cod skin collagen peptide mixed peptide from Alaska cod skin by usinga protease enzymolysis method; then carrying out ultrafiltration treatment, sephadex chromatographic column rough separation and high performance liquid chromatography separation in sequence; whereinthe sephadex chromatographic column is formed by connecting sephadex resin Sephadex G-25 and sephadex resin Sephadex G-50 in series. The activity test shows that the three kinds of cod skin oligopeptides have alpha-glucosidase inhibitory activity, can assist in reducing blood sugar, and can be used for preparing medicine for treating type II diabetes.

Description

technical field [0001] The invention relates to the field of separation, purification and application of fish skin oligopeptides, in particular to the application of Alaska pollock skin oligopeptides and their separation and purification method in the preparation of α-glucosidase inhibitors and anti-type Ⅱ diabetes medicines. Background technique [0002] Alaskan pollock. The scientific name is Theragra chalcogramma, which lives in the northern Atlantic Ocean. It is a cold-water deep-sea fish with tender meat and light meat taste. Due to its special long-term growth environment (low temperature, high pressure), its fish protein (fish meat, fish skin, etc.) The amino acid composition and amino acid sequence may be different from other shallow sea fish and freshwater fish. [0003] Cod skin collagen peptide is a mixture of proteins, polypeptides, oligopeptides and amino acids (the content is mainly oligopeptides) prepared from cod skin by enzymatic method. The oligopeptides ...

Claims

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Application Information

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IPC IPC(8): C07K5/087C07K7/06C12P21/06C07K1/34C07K1/16C07K1/14C07K1/36A61K38/08A61P3/10
CPCC07K5/0812C12P21/06A61P3/10A61K38/00C07K7/06
Inventor 贾建萍包国良李青青
Owner ZHEJIANG ACAD OF MEDICAL SCI
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