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Method for breeding enramycin strain by induced mutation of cabicidin streptomycete with ion beam

A fungicide, enramycin technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., to achieve the effects of simple equipment, great promotion value, and good mutagenesis effect

Inactive Publication Date: 2013-04-03
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no related patents and literature reports on the selection of ion beam implantation in the production of enramycin strains.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] At a room temperature of 25°C, the strain FYFJ03 was inoculated into a liquid fermentation medium, cultured on a shaker at 28°C, 200-220 rpm for 160 hours, and the production of enramycin in the fermentation broth was detected, and the fermentation unit was 910 U / mL.

Embodiment 2

[0027] At room temperature of 25°C, the slant spores of strain FYFJ03 were taken to make 10 5 ~10 7 spores / mL spore suspension, spread evenly on a sterile plate, blow dry under sterile conditions, use a nitrogen ion beam with an energy of 10Kev, and a dose of 1.5×10 14 ions / cm 2 Pulse injection was performed under the condition of 5 After doubling, spread it on a solid medium, culture it at 28°C for 9 days, select a single strain that grows well and inoculate it in a liquid fermentation medium, culture it on a shaker at 28°C, 200-220rpm for 144 hours, and detect enramycin in the fermentation broth Yield, the fungicide Streptomyces mutant strains with stable genetic traits of high enramycin production were selected. The positive mutation rate of the mutant strain was 19%. The average fermentation unit is 1146.6U / mL, an average increase of 26% compared with the original strain.

Embodiment 3

[0029] At room temperature of 25°C, the slant spores of strain FYFJ03 were taken to make 10 5 ~10 7 spores / mL spore suspension, spread evenly on a sterile plate, blow dry under sterile conditions, use a nitrogen ion beam with an energy of 10Kev, and a dose of 3×10 14 ions / cm 2 Pulse injection was performed under the condition of 5 After doubling, spread it on a solid medium, culture it at 28°C for 10 days, select a single strain that grows well and inoculate it in a liquid fermentation medium, culture it on a shaker at 28°C, 200-220rpm for 156 hours, and detect enramycin in the fermentation broth Yield, the fungicide Streptomyces mutant strains with stable genetic traits of high enramycin production were selected. The positive mutation rate of the mutant strain was 15%. The average fermentation unit is 1192.1U / mL, an average increase of 31% compared with the original strain.

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Abstract

The invention provides a method for breeding an enramycin strain by induced mutation of cabicidin streptomycete with an ion beam. The method comprises the following steps of: (1) uniformly coating a spore suspension prepared from cabicidin streptomycete slant spores on a sterile plate at room temperature; (2) carrying out pulse injection on the spore suspension by using a nitrogen ion beam; (3) diluting the spore suspension subjected to treatment in the step (2), and then coating the diluted spore suspension in a solid culture medium for culture; and (4) selecting a single strain with good growth conditions, inoculating the single strain in a liquid fermentation culture medium for culture, and selecting a high-yield strain with stable genetic characters, wherein the high-yield strain is the enramycin strain. The method provided by the invention has the advantages of simple equipment, feasibility and safety in operation by adoption of the ion beam for induced mutation, and has an induced mutation effect far better than that of the traditional physical chemical induced mutation method; and the cabicidin streptomycete obtained by induced mutation based on the method is improved by 20-40% in fermentation unit compared with that of an original strain.

Description

technical field [0001] The invention relates to the technical field of microbial mutagenesis, in particular to a method for breeding enramycin strains by ion beam mutagenesis of fungicide streptomyces. Background technique [0002] Enramycin (Enramycin), also known as Enlaimycin, Enmycin, and Duramycin, is a cyclic polypeptide antibiotic composed of 13 different types of 17 amino acid molecules and fatty acid molecules, and the amino acids form a ring Polypeptides, fatty acid molecules connected to the terminal aspartic acid, are divided into enramycin A and enramycin B according to the difference of terminal fatty acid molecules, and enramycin is a mixture of these two components. The drug was developed by Takeda Pharmaceutical Co., Ltd. in Japan in 1966, officially registered in Japan in 1974, and has since been registered and widely used in many countries. In 1993, the Ministry of Agriculture of my country approved the registration of the drug in my country. Enramycin c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N1/20C12R1/465
Inventor 李荣杰尚海涛郑辉杨金环
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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