Vibrio FC509 strain and culturing method and application thereof

A technology of Vibrio and bacterial strains, applied in the field of microorganisms, to achieve the effect of broad application prospects

Active Publication Date: 2014-07-16
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few highly active glycosaminoglycan-degrading enzymes with application value at present, so it is of great significance to isolate and identify new high-efficiency and stable glycosaminoglycan-degrading enzymes from new glycosaminoglycan-degrading bacteria

Method used

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  • Vibrio FC509 strain and culturing method and application thereof
  • Vibrio FC509 strain and culturing method and application thereof
  • Vibrio FC509 strain and culturing method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1, the acquisition of Vibrio sp. FC509 strain

[0037] A strain of Vibrio sp. FC509 was deposited in the General Microbiology Center of China Committee for the Collection of Microbial Cultures on March 12, 2014, with the preservation number CGMCC NO.8913, address: No. 1, Beichen West Road, Chaoyang District, Beijing 3 Institute of Microbiology, Chinese Academy of Sciences.

[0038] Take the soil leachate, add 1 mL of the supernatant to 9 mL of sterile water, dilute to a suitable concentration gradient of 10-1, 10-2, 10-3, 10-4, 10-5, 10-6, and then pass through the conventional Dilution method or streaking method to inoculate and spread on the only carbon source separation medium, culture the bacteria at a constant temperature of 30°C for 1 day, count the colonies, then select colonies with obvious differences in colony morphology, and repeatedly streak and inoculate on the corresponding complete nutrition on the agar plate until a single colony was obtained af...

Embodiment 2

[0044] The cultivation method of Vibrio sp. FC509 bacterial strain, the steps are as follows:

[0045] (1) Take Vibrio sp. FC509 strain and inoculate it into the liquid medium, and cultivate it on a shaking table for 10 hours at a temperature of 28-30°C and a rotation speed of 200rpm to obtain a seed solution;

[0046] (2) Take the seed solution prepared in step (1), inoculate it in the liquid medium at a volume percentage of 5-10%, and expand the culture for 2 hours at a temperature of 25-30°C and a dissolved oxygen of 30%. , to obtain Vibrio sp. FC509 bacteria liquid.

[0047] Preferably according to the present invention, the components per liter of the liquid culture medium are as follows:

[0048] Chondroitin Sulfate 5g, Tryptone 10g, Yeast Extract 5g, NaCl 30g, KH 2 PO 4 3g,K 2 HPO 4 7g, (NH 4 ) 2 SO 4 2g, MgSO 4 ·7H 2 05mg, FeSO 4 ·7H 2 02mg, pH 7.2.

Embodiment 3

[0050] The application of Vibrio sp. FC509 strain in the preparation of glycosaminoglycan lyase, the steps are as follows:

[0051] (1) Take Vibrio sp. FC509 strain and inoculate it into the liquid medium, and cultivate it on a shaker for 16 hours at a temperature of 25-28°C and a rotation speed of 300rpm to obtain a seed solution;

[0052] (2) Take the seed solution prepared in step (1), inoculate it in the liquid medium at a volume percentage of 5-10%, and expand the culture for 5 hours at a temperature of 25-30°C and a dissolved oxygen of 25%. , adding chondroitin sulfate so that the mass concentration of chondroitin sulfate is 0.01-0.02%, and continuing to cultivate for 72 hours to obtain Vibrio sp. FC509 strain fermentation broth;

[0053] (3) Take the fermented broth of Vibrio sp. FC509 strain obtained in step (3), separate the solid from the liquid, take the liquid, add ammonium sulfate to make the mass concentration of ammonium sulfate 80%, collect the precipitate, and...

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Abstract

The invention relates to a Vibrio FC509 strain and a culturing method and application thereof. The Vibrio FC509 strain is collected in the China General Microbiological Culture Collection Center (CGMCC) on 12, March, 2014, with a collection number of CGMCC NO.8913 and an address of Institute of Microbiology Chinese Academy of Sciences NO.1 West Beichen Road, Chaoyang District, Beijing. The Vibrio sp. FC509 strain obtained by first-time separation is capable of preparing glycosaminoglycan lyase which has a specific activity of 110,000U / mg to hyaluronic acid and specific activity of 45,000U / mg to chondroitin sulfate; enzyme activity is tens to hundreds times that of the known commercialized glycosaminoglycan lyase, can be applied to fields such as medicines, cosmetics and the like, and is wide in application prospect.

Description

technical field [0001] The invention relates to a strain of Vibrio FC509 and its culture method and application, belonging to the technical field of microorganisms. Background technique [0002] Glycosaminoglycans (Proteoglycans, GAGs), also known as mucopolysaccharides, are linear polysaccharides composed of repeating disaccharide units. Mainly include hyaluronic acid (Hyaluronic Acid, HA), heparin / heparan sulfate (Hep / HS), chondroitin sulfate / dermatan sulfate (Chondroitin Sulfate / Dermatan Sulfate, CS / DS), keratan sulfate (Keratan Sulfate, KS). Except that the disaccharide unit of keratan sulfate is composed of neutral D-galactose and N-acetylglucosamine, the disaccharide units of other glycosaminoglycans are composed of hexuronic acid (D-glucuronic acid / L -iduronic acid) and hexosamine (N-acetylglucosamine / N-acetylgalactosamine). Among them, the structure of hyaluronic acid is relatively simple, and the disaccharide units composed of D-glucuronic acid and N-acetylglucos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12R1/63
Inventor 李福川韩文君王文爽赵梅
Owner SHANDONG UNIV
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