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Streptomyces clavulcane, application, fermentation medium and preparation method of clavulanic acid

A technology of Streptomyces clavulanicus and fermentation medium, which is applied in the biological field and can solve the problems of low clavulanic acid production and inability to meet the needs of industrial fermentation

Active Publication Date: 2022-05-06
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the clavulanic acid production of wild-type Streptomyces clavulanic acid is very low, which is far from meeting the needs of industrial fermentation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Screening of Streptomyces clavuligerus SIIA19-77# and preparation of clavulanic acid

[0036] 1. Streptomyces clavuligerus SIIA0855# is obtained from Streptomyces clavuligerus SIIA0198# (provided by Sichuan Antimicrobial Industry Research Microorganism Resource Center) through multiple rounds of mutagenesis screening, and its clavulanic acid The fermentation unit is 4100μg / mL~4200μg / mL.

[0037] 2, (1) select the clavulanic acid producing streptomyces SIIA0855# slant bacterial strain that grows well, wash down spore with physiological saline, filter through cotton, make 10 6 Individual / mL spore suspension;

[0038] (2) Use a pipette gun to draw 10 μL of the spore suspension in (1) onto a glass slide with a diameter of 1 cm, and place it in a room-temperature plasma induction chamber with helium as the working gas, a power of 120 W, and a ventilation rate of 10 SLM. In the mutagenesis system, the distance between the plasma emission source and the sample was 2mm, and t...

Embodiment 2

[0053] The only difference with Example 1 is:

[0054] In terms of mass percentage, the composition of the separation medium is: 1% soluble starch, 0.0512% anhydrous magnesium sulfate, 0.05% potassium nitrate, 0.025% ferrous sulfate, 0.15% sodium chloride, 0.02% dipotassium hydrogen phosphate, threonine 10%, agar 1.5%, pH7.0.

[0055] Inoculate the Streptomyces clavuligerus SIIA19-77# isolated in step (6) on the slant medium, culture at 28°C for 6-8 days, and obtain the slant strain;

[0056] Inoculate the slant strain into the seed culture medium at 28°C, at a rotation speed of 220r / min, and cultivate for 32h to obtain a seed solution;

[0057] The seed liquid was inoculated into the fermentation medium at 28°C with a rotation speed of 220r / min at an inoculum amount of 10% (V / V), cultured for 96 hours, and the fermentation liquid was collected to obtain clavulanic acid;

[0058] In terms of mass percentage, the slant culture medium consists of: 2% soluble starch, 0.0244% an...

Embodiment 3

[0063] The only difference with Example 1 is:

[0064] In terms of mass percentage, the composition of the separation medium is: 2% soluble starch, 0.0244% anhydrous magnesium sulfate, 0.1% potassium nitrate, 0.001% ferrous sulfate, 0.05% sodium chloride, 0.05% dipotassium hydrogen phosphate, threonine 5%, agar 1.5%, pH7.0.

[0065] Inoculate the Streptomyces clavuligerus SIIA19-77# isolated in step (6) on the slant medium, culture at 28°C for 6-8 days, and obtain the slant strain;

[0066] Inoculate the slant strain into the seed culture medium at 28°C, at a rotation speed of 220r / min, and cultivate for 32h to obtain a seed solution;

[0067] The seed solution was inoculated into the fermentation medium at 28°C with a rotation speed of 220r / min at an inoculum amount of 15% (V / V), cultured for 96 hours, and the fermentation broth was collected to obtain clavulanic acid;

[0068] In terms of mass percentage, the slant culture medium consists of: 2% soluble starch, 0.0244% anh...

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PUM

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Abstract

The invention discloses Streptomyces clavuligerus, its application, a fermentation medium and a preparation method of clavulanic acid; the Streptomyces clavuligerus is preserved in the General Microorganism Center of China Committee for the Collection of Microorganisms, and the preservation number is CGMCC No.19076; Disclosed is the application of Streptomyces clavulanic acid in the production of clavulanic acid; the preparation method of clavulanic acid is disclosed comprising: fermenting the Streptomyces clavulanic acid to obtain clavulanic acid; the Streptomyces clavulanicus has good genetic stability, and the output of clavulanic acid is large , high potency.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a streptomyces clavulcane, its application, a fermentation medium and a preparation method of clavulanic acid. Background technique [0002] The abuse of β-lactam antibiotics such as penicillins and cephalosporins causes some pathogenic bacteria to produce β-lactamase, which degrades these antibiotics and produces drug resistance. Clavulanic acid is a natural β-lactamase inhibitor produced by Streptomyces clavuligerus, which can synergistically enhance the antibacterial activity of penicillins and cephalosporins against β-lactamase-resistant bacteria. Greatly improve the sensitivity of pathogenic bacteria to β-lactam antibiotics. Augmentin for clinical use is a compatible preparation of clavulanic acid and amoxicillin, and timentin is a compatible preparation of ticarcillin sodium and clavulanic acid potassium, and clavulanic acid is widely used in clinical practice. [0003] At pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/18C12R1/465
CPCC12N1/20C12P17/188C12R2001/465C12N1/205
Inventor 俞岩青王昆蓉田敏雷叶明
Owner CHENGDU UNIV
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