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Method for separating alpha-glucosidase inhibitor from laver enzymolysis product

A technology of glucosidase and enzymatic hydrolysis products, which is applied in the field of separation and purification of biological products, can solve the problems of high price and scarce sources, achieve the effects of small molecular weight, prevent resource waste, and high stability

Active Publication Date: 2013-12-25
NANTONG CHITSURU FOODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The domestic demand for α-glucosidase inhibitors is huge, mainly relying on imports, but the sources are scarce and expensive, which brings heavy economic pressure to the majority of diabetic patients

Method used

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  • Method for separating alpha-glucosidase inhibitor from laver enzymolysis product
  • Method for separating alpha-glucosidase inhibitor from laver enzymolysis product
  • Method for separating alpha-glucosidase inhibitor from laver enzymolysis product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] With 3 g of dried Porphyra striatus powder dried to constant weight as substrate, pH7.0 Na 2 HPO 4 -KH 2 PO 4 The buffer was used as the medium, the ratio of substrate to buffer was 1:33, and heat treatment was carried out in a boiling water bath for 15 min. Add neutral protease (E / S=8×10 4 U / g ), hydrolyzed at 50°C for 7 h, inactivated the enzyme in boiling water bath for 15 min; adjusted the pH to 8.5, added solid aminopeptidase ( E / S=28 U / g ) and alkaline protease ( E / S=8×10 4 U / g ), hydrolyze at 50°C for 3 h, inactivate the enzyme in a boiling water bath for 15 min, and add ethanol to a final concentration of 60%. Let it stand overnight at room temperature, centrifuge at 8000 r / min for 10 min, and the supernatant obtained is a mixture containing α-glucosidase inhibitor. The inhibitor-containing laver supernatant mixture was decolorized by activated carbon, and then separated and purified by 1 mL prepacked column SP Sepharose High Performance cation exchange c...

Embodiment 2

[0038] With 4 g of Porphyra zebra powder dried to constant weight as substrate, pH7.0 Na 2 HPO 4 -KH 2 PO 4 The buffer was used as the medium, the ratio of substrate to buffer was 1:25, and heat treatment was carried out in a boiling water bath for 15 min. Add neutral protease (E / S=8×10 4 U / g ), hydrolyzed at 50°C for 7 h, inactivated the enzyme in a boiling water bath for 15 min, adjusted the pH to 8.5, added solid aminopeptidase ( E / S=28 U / g ) and alkaline protease ( E / S=8×10 4 U / g ), hydrolyze at 50°C for 3 h, inactivate the enzyme in a boiling water bath for 15 min, and add ethanol to a final concentration of 60%. After standing overnight at room temperature, centrifuge at 8000 r / min for 10 min, and the obtained supernatant is a mixture containing inhibitors. The inhibitor-containing laver supernatant mixture was decolorized by activated carbon, and then separated and purified by 1 mL prepacked column SP Sepharose High Performance cation exchange chromatography. Eq...

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Abstract

The invention discloses a method for separating an alpha-glucosidase inhibitor from a laver enzymolysis product, which belongs to the technical field of separation and purification of biological products. The invention relates to a method for compounding porphyra yezoensis to obtain a rough alpha-glucosidase inhibitor product, wherein the enzymolysis is controllable and the cost is low. The rough product is separated and purified through SPSepharoseHighPerformance cation exchange chromatography, SephadexG-10 gel chromatography and Mono Q anion exchange chromatography, and the purity and main composition of an LGI (lactate glucose index) part with high inhibitory activity on alpha-glucosidase are analyzed through reversed phase high performance liquid chromatography and ultra-high performance liquid chromatography-mass spectrography. The IC 50 value of LGI on the alpha-glucosidase is 97.63mu g / ml, and the temperature and pH stability are good; the inhibitory activity is kept to be higher than 80% in 80 DEG C water bath for 7 h, and the inhibitory activity is still kept to be higher than 87% after being maintained in a buffer solution with pH of 3-12 at 50 DEG C for 2 h. The LGI is small in molecular weight, good in stability, low in cost and safe to use, thereby having a quite wide application prospect in such fields as medicine, food, etc.

Description

technical field [0001] The invention discloses a method for separating α-glucosidase inhibitors from laver enzymatic hydrolyzate, which belongs to the technical field of separation and purification of biological products. Background technique [0002] Diabetes is a common endocrine metabolic disease characterized by hyperglycemia caused by living environment and genetic factors. According to the statistics of the World Health Organization (WTO), more than 350 million people around the world are suffering from diabetes. Diabetics in my country account for about one-third of the world's diabetic population, and the number is expected to double by 2025. Diabetes and its complications have become the third biggest killer of human after tumor and cardiovascular disease. The medical profession usually divides it into two categories: type 1 diabetes and type 2 diabetes. The pathogenesis of diabetes has not been fully elucidated, but the main reason can be attributed to the abnorm...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/18C07K1/16
Inventor 田亚平周锋刘冬冬
Owner NANTONG CHITSURU FOODS
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