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Streptomyces fungicidious mutant strain and application thereof

A technology for mutant strains and fungicides is applied in the field of strains for preparing polypeptide antibiotics, which can solve the problems of unfavorable metabolic pathways, low efficiency, unsatisfactory results, etc. Reliable effects of screening methods

Active Publication Date: 2012-05-23
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous screening was limited to screening wild-type strains from the environment, which not only involved heavy workload, low efficiency, but also unsatisfactory results
This may be due to the fact that the metabolic pathway of the wild-type strain is not conducive to the direction of enramycin synthesis, or the amount of synthetic precursors is insufficient

Method used

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  • Streptomyces fungicidious mutant strain and application thereof
  • Streptomyces fungicidious mutant strain and application thereof
  • Streptomyces fungicidious mutant strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Obtaining of anti-α-amino-β-hydroxyvaleric acid mutant strain

[0020] 1. Strain treatment: inoculate the fungicide Streptomyces fungicidious (ATCC 27432) on oat flour agar-1 (oat flour 20g / L, FeSO 4 ·7H 2 O0.1g / L, MnCl 2 4H 2 O 0.1g / L, ZnSO 4 ·7H 2 O 0.1g / L, agar 15g / L) slant medium, 28 ℃ constant temperature culture. Pick an appropriate amount of single bacterium colony formed by streaking with a sterilized toothpick and inoculate it in liquid oat flour medium (oat flour 20g / L, FeSO 4 ·7H 2 O 0.1g / L, MnCl 2 4H 2 O 0.1g / L, ZnSO 4 ·7H 2 (20.1g / L), under the condition that temperature is 28 ℃, rotating speed is 200rpm, culture thalline to logarithmic growth phase, get the bacterial strain of this growth phase and do chemical mutagenesis experiment.

[0021] 2. Strain mutation and screening: take the above-mentioned bacterial solution and centrifuge at 5000rpm for 10 minutes to collect the bacterial cells, add physiological saline to form a suspension...

Embodiment 2

[0022] The screening of embodiment 2 enramycin high-yielding bacterial strains

[0023] 1. Activation of Staphylococcus aureus: Streak the Staphylococcus aureus preserved on the slant on the nutrient gravy (peptone 10g / L, beef extract 3g / L, NaCl 5g / L) agar plate, culture at 37°C 16h. Use a sterilized toothpick to pick an appropriate amount of the single colony formed by activation and inoculate it in liquid nutrient gravy agar medium, shake and culture it at 37°C and 200rpm for 14-16h, and cultivate the OD of the bacteria. 600 When the logarithmic growth phase reaches 1.0-1.5, draw 120 μL and spread it on the nutrient broth agar plate, cultivate at 37°C, and set aside.

[0024] 2. Screening of enramycin-producing bacteria: get the single colony of the anti-α-amino-β-hydroxyvaleric acid mutant strain in Example 1 in liquid oat flour medium (substrate composition is the same as described in Example 1) Cultured in medium for 5-7 days, soaked with sterile filter paper with a dia...

Embodiment 3

[0025] The fermentation of embodiment 3 enramycin high-yield bacterial strains

[0026] Get the bacterial strain screened in Example 2 as the fermenting strain, take the starting strain as the contrast, inoculate respectively in the fermentation medium and ferment, the components of the fermentation medium are corn flour 35g / L, corn steep liquor 25g / L, soybean cake powder 34.0 g / L, soluble starch 60g / L, glucose 20g / L, calcium carbonate 30g / L, FeSO 4 ·7H 2 O 2g / L, MnCl 2 4H 2 O 2g / L, ZnSO 4 ·7H 2 O 0.1g / L. The temperature is 28° C., 200 rpm, and the fermentation time is 7 days.

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Abstract

The invention provides a streptomyces fungicidious mutant strain FYFJ03 of which the collection number is CGMCC No.4113, the collection institution is China General Microbiological Culture Collection Center, and the collection date is August 25th, 2010. The invention also provides the application of the strain in the production of enramycin. In the mutant strain provided by the invention, threonine can be accumulated and the content of primer threonine for synthesizing the enramycin can be increased, so that the production level of the enramycin is improved.

Description

technical field [0001] The invention relates to a bacterial strain for preparing polypeptide antibiotics, in particular to a fungicide Streptomyces mutant strain and its application in the production of enramycin. Background technique [0002] Enramycin (Enramycin), also known as Enlaimycin, Anlemycin, and Duramycin, is a cyclic polypeptide antibiotic composed of 13 different types of 17 amino acid molecules and fatty acid molecules. Amino acids form a ring The fatty acid molecule is connected to the aspartic acid at the end. According to the difference of the terminal fatty acid molecule, it is divided into enramycin A and enramycin B. Enramycin is a mixture of these two components. [0003] Enramycin has a strong inhibitory effect on Gram-positive bacteria, mainly hindering the synthesis of bacterial cell walls. Sensitive bacteria include various Gram-positive cocci such as Staphylococcus aureus, Bacillus subtilis, Bacillus anthracis, Clostridium tetani, Clostridium botul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/01C12P21/04C12R1/465
Inventor 徐斌崔刚
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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