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Streptomyces lavendulae and method and application thereof for preparing alpha-glucosidase inhibitor

A technology of Streptomyces lilacinus and glucosidase is applied in the preparation of α-glucosidase inhibitor, the application field of α-glucosidase inhibitor in the treatment of diabetes, and the formation of postprandial hyperglycemia can be improved and reduced. Effects of postprandial blood sugar levels

Inactive Publication Date: 2016-03-09
GUANGXI NANNING ZHITIAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The above patents are all about the application of chemically synthesized drugs in the preparation of α-glucosidase inhibitors. At present, there is no document disclosing Streptomyces lilacinus and its method and application for the preparation of α-glucosidase inhibitors.

Method used

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  • Streptomyces lavendulae and method and application thereof for preparing alpha-glucosidase inhibitor
  • Streptomyces lavendulae and method and application thereof for preparing alpha-glucosidase inhibitor
  • Streptomyces lavendulae and method and application thereof for preparing alpha-glucosidase inhibitor

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Experimental program
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Effect test

Embodiment 1

[0047] Embodiment 1: Isolation and screening of Streptomyces lilacinus gray Streptomyces UN-8

[0048] (1) Preparation of bacterial suspension: Weigh 5 g of soil samples into a 250 mL Erlenmeyer flask filled with 45 mL of sterile saline, and stir magnetically for 30 minutes at room temperature to prepare bacterial suspension.

[0049] (2) Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 4 , 10 5 , 10 6 100 μL of the bacterial suspension was spread on the solid separation medium plate; after culturing at 28°C for 5 days, single colonies were picked according to the colony shape and size of the actinomycetes and the color of the pigment produced.

[0050] (3) Streak purification: use a flat and smooth inoculation loop to continuously streak and purify the obtained single colony on the Gaoshi No. 1 solid medium plate under asept...

Embodiment 2

[0058] Embodiment 2: the culture characteristic of streptomyces lilacinus griseus UN-8

[0059] Streptomyces lilacinus griseus UN-8 was inoculated in a 250mL Erlenmeyer flask equipped with 50mL carbon-free liquid medium, and the liquid medium added with 2.5g / L and 10g / L glucose was used as a control group respectively, and the shaking table speed was 160rpm. After culturing at 28°C for 7 hours, the growth of the strain was judged by the dry weight method or turbidimetric method. The results showed that the growth of Streptomyces lilacaceae UN-8 in the carbon-free liquid medium and 2.5g / L glucose liquid medium was good. And it is better than the growth of Streptomyces lilacinus UN-8 in 10g / L glucose liquid medium.

[0060] Inoculate Streptomyces lavenderus UN-8 in a 250mL Erlenmeyer flask filled with 50mL nitrogen-free liquid medium, add 1g / L yeast extract and 1g / LKNO 3 The liquid culture medium was used as the control group, the shaker speed was 160rpm, and after culturing ...

Embodiment 3

[0063] Embodiment 3: the method for preparing α-glucosidase inhibitor by Streptomyces lilacaceae UN-8

[0064] (1) Fermentation: Use a flat and smooth inoculation loop to pick a lawn with a length of 3 cm on the inclined surface and insert it into a 500 mL conical flask containing 100 mL of liquid seed medium. Put 10% of the inoculum into a 3L Erlenmeyer flask containing 800mL of liquid fermentation medium, shake the table at a speed of 160rpm, cultivate at 30°C for 3 days, collect the fermentation broth of 5 Erlenmeyer flasks, about 4L, and use it for α-glucosidase Separation and extraction of inhibitors;

[0065] (2) Pretreatment of fermentation broth: Centrifuge the fermentation broth at 8000rpm and 4°C for 15 minutes to remove bacteria and insoluble particles, then filter the supernatant through an ultrafiltration system with a molecular weight cut-off of 5000Da, and the average membrane flux is 30LMH, remove the residual soluble starch, protein and part of the pigment ...

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Abstract

The invention discloses streptomyces lavendulae of which the classification is named as Streptomyces lavendulae UN-8, the preservation number is CCTCC NO: M 2015512, the preservation date is September 6th, 2015, and the preservation unit is China Center for Type Culture Collection. The Streptomyces lavendulae UN-8 has application in the aspects of fermenting and preparing alpha- glucosidase inhibitor. A certain amount of fermentation liquor of the alpha-glucosidase inhibitor is accumulated through strain fermentation, bactofugation is conducted, soluble starch, protein and part pigment are removed through ultrafiltration, decoloration is conducted through activated carbon, nanofiltration concentration is conducted, most inorganic salt micromolecules are removed, adsorption and desorption are conducted through resin, decompression drying is conducted, and an alpha-glucosidase inhibitor refined product is obtained. The alpha-glucosidase inhibitor can strongly inhibit alpha-glucosidase, and IC50 of the alpha-glucosidase inhibitor is 321 times that of commercially available alpha-glucosidase inhibitor acarbose; the obvious improvement action on forming of postprandial hyperglycemia inside an animal is achieved, the effect is equal to that of the acarbose, and the alpha-glucosidase inhibitor can be used for preparing drugs for treating diabetes or functional foods.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to Streptomyces lilacaceae and a method for preparing an α-glucosidase inhibitor, as well as the application of the α-glucosidase inhibitor in treating diabetes. Background technique [0002] Diabetes mellitus is a comprehensive metabolic disease with persistent high blood sugar as the main biochemical feature. Clinically, it is mainly divided into type I diabetes and type II diabetes. In my country, type II diabetes is the majority. The clinical symptoms are abnormally high glucose concentration in blood and urine. Patients have obvious symptoms of polydipsia, polyphagia, polyuria, and weight loss, accompanied by symptoms of fatigue and weakness. With the exacerbation of the disease, persistent high blood sugar can cause many complications, endanger health, and cause disability or death in severe cases. With the continuous improvement of people's living stan...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/06A61K35/74A61P3/10A23L33/00C12R1/56
Inventor 梁智群陈桂光曾伟黄熙李娟
Owner GUANGXI NANNING ZHITIAN BIOTECH
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