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Streptomyces clavuligerus strain with high production of clavulanic acid and stress breeding method and application thereof

A technology of Streptomyces clavulcane and clavulanic acid, applied in the direction of using microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problem of unclear direction of breeding, long breeding cycle, and low breeding efficiency problems such as high potency, large yield and good genetic stability

Inactive Publication Date: 2019-06-21
HEZE RUIZHI TECH DEV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to solve the problems of the prior art, such as the unclear direction of positive and negative mutation breeding, long breeding cycle, low breeding efficiency, and high investment in equipment, etc., and make up for the deficiencies of the prior art, the present invention provides a high-yield strain of Streptomyces clavulicularis, which Uses and stress breeding methods

Method used

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  • Streptomyces clavuligerus strain with high production of clavulanic acid and stress breeding method and application thereof
  • Streptomyces clavuligerus strain with high production of clavulanic acid and stress breeding method and application thereof
  • Streptomyces clavuligerus strain with high production of clavulanic acid and stress breeding method and application thereof

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Effect test

Embodiment 1

[0050] 1.1 Experimental materials:

[0051] Streptomyces clavulicularis B-7; ATCC 27064;

[0052] Klebsiella: Klebsiella ATCC 31488;

[0053] Streptomyces clavulicularis isolation medium: yeast powder 0.4%, glucose 0.4%, malt powder 1%, agar powder 2.0%, pH adjustment 8.2;

[0054] The fermentation medium is: soybean protein 5.2%, dextrin 0.96%, MOPS 1.05%, glycerol 1.5%, KH 2 PO 4 0.12%, CaCl 2 0.01%, FeCl 3 0.003%, ZnSO 4 ·7H 2 O 0.0005%, MgSO 4 0.0005%, CuCl 2 0.0005%, soybean oil 2.2%, pH modulation 7.0.

[0055] 1.2 Experimental process:

[0056] The stress breeding method of the Streptomyces clavulanicus bacterial strain B-77 of high-yield clavulanic acid, comprises steps:

[0057] 1) Strain activation: Take out the frozen Streptomyces clavicularis B-7 from the -80°C refrigerator, let it thaw at room temperature, use a pipette gun to take 10uL and drop it on the slant of Streptomyces clavicularis isolation medium, and use "Z" to draw a line French line. Cul...

Embodiment 2

[0070] Example 2 Studies on the Genetic Stability of Streptomyces clavulatus Strain B-77

[0071] 1.1 Experimental materials:

[0072] Mutant strain: Streptomyces clavulicularis strain B-77;

[0073] Slant medium: yeast powder 2%, glucose 0.4%, malt powder 1%, agar powder 2.0%, pH adjustment 8.2;

[0074] Fermentation medium: soybean protein 5.2%, dextrin 0.96%, MOPS 1.05%, glycerin 1.5%, KH 2 PO 4 0.12%, CaCl 2 0.01%, FeCl 3 0.003%, ZnSO 4 ·7H 2 O 0.0005%, MgSO 4 0.0005%, CuCl 2 0.0005%, soybean oil 2.2%, pH modulation 7.0.

[0075] 1.2 Experimental process

[0076] 1. Inoculate the mutant strain Streptomyces clavulus strain B-77 into the sterilized slant medium, carry out subcultivation, pass 6 generations continuously, and store the cultured strain in a refrigerator at 4°C;

[0077] 2. Inoculate the cultured slant into the fermentation medium of the Erlenmeyer flask, and culture it on a shaker at 25° C. and 250 rpm for 96 hours, measure the potency of clavulanic...

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Abstract

The invention discloses a Streptomyces clavuligerus strain with high production of clavulanic acid and a stress breeding method and application thereof, and belongs to the technical field of biology.The Streptomyces clavuligerus strain with high production of clavulanic acid disclosed herein is collected in China Center for Type Culture Collection on February 25th, 2019 under CCTCC No. 2019110. The Streptomyces clavuligerus strain B-77 with high production of clavulanic acid is cultured by adding stress materials (penicillin and Klebsiella) into a culture medium under suitable adding mode ofthe stress materials, suitable adding time, suitable culture conditions, suitable separate culture process of the strain and the like. The strain acquired via the method herein has good genetic stability; the ability of the strain to produce clavulanic acid is improved significantly; the yield of clavulanic acid is high, and titer is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a strain of Streptomyces clavulanicum with high yield of clavulanic acid, its stress breeding method and its application. Background technique [0002] Clavulanic Acid (CA) is a β-lactamase inhibitor isolated and purified from the fermentation broth of Streptomyces clavuligerus. Its molecule is a fused bicyclic β-lactam structure, which is replaced by an oxygen atom The sulfur atom in the thiazole ring of penicillin and cephalosporin has been removed, and it has the necessary 3R, 5R stereochemical structure as a β-lactamase inhibitor. Clavulanic acid itself has weak antibacterial activity, but it is a potent, broad-spectrum and irreversible β-lactamase inhibitor. Produces β-lactamases produced by drug-resistant Gram-positive and Gram-negative bacteria, especially Klebsiella pneumoniae, Proteus mirabilis and Staphylococcus aureus, both in vitro and in vivo Strong and irreversible co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/36C12P17/18C12R1/465C12R1/22
Inventor 周汝德董庆茹李振森吴桂苹
Owner HEZE RUIZHI TECH DEV
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