A kind of high-yielding strain of avilamycin and its breeding method

A technology of avelamycin and high-yield strains, applied in the field of microbial breeding, to achieve stable performance, high yield, and reduce workload

Active Publication Date: 2020-01-03
杭州皇冠农业生物工程技术研究中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The productivity of the high-yield avelamycin-producing bacteria published in this paper still needs to be improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A kind of avelamycin high-yield strain breeding method, concrete steps:

[0022] 1) ARTP and the first time 137 Csy mutagenesis: the original strain 11-10 was activated and cultured to prepare single spore suspension, ARTP and 137 For Csy mutagenesis, the irradiation time of ARTP is 170-210s. The higher lethality rate is beneficial to eliminate parents and screen mutants, and the plasma mutagenesis time should not be too long. The agar block method was used for primary screening, shake flask fermentation for re-screening, and the potency was measured to obtain mutagenic high-yield strains B-1, F-2 and F-28, which were stored in freeze-dried tubes. Plasma mutagenesis uses a plasma source capable of generating a plasma jet at atmospheric pressure with a temperature between 25-40°C and a high concentration of reactive ions. The active particles in the plasma can change the permeability of the cell walls and membranes of microorganisms, causing corresponding gene damage, ...

Embodiment 2

[0033] Streptomyces viridochromogenes 11-10, with a potency of 710 mg / L, provided by the laboratory of Zhejiang Gongshang University.

[0034] Test bacteria: Micrococcus luteus (Microccus luteus) 10209, from China Industrial Microbiology Culture Collection Center.

[0035] A kind of avelamycin high-yield strain breeding method, concrete steps:

[0036] 1) ARTP and the first time 137 Csy mutagenesis: the original strain 11-10 was activated and cultured to prepare single spore suspension, ARTP and 137 For Csy mutagenesis, the irradiation time of ARTP is 200s, and the higher lethality rate is beneficial to eliminate parents and screen mutants, and the plasma mutagenesis time should not be too long. The agar block method was used for primary screening, shake flask fermentation for re-screening, and the potency was measured to obtain mutagenic high-yield strains B-1, F-2 and F-28, which were stored in freeze-dried tubes. Plasma mutagenesis uses a plasma source capable of generat...

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Abstract

The invention provides a kind of avelamycin high-yield bacterial strain breeding method, adopts ARTP and three times 137 Csy mutagenesis mutated Streptomyces and used 2 protoplast fusions for gene recombination. The beneficial effect is: after four rounds of mutagenesis and two rounds of protoplast fusion, the fusion high-yield strain G2-31 was obtained, and its avelamycin titer reached 2842.57 mg / L, which was 4 times that of the original strain 11-10. Passage test, strain state is stable, showing high potential for industrial application.

Description

technical field [0001] The invention relates to the technical field of microbial breeding, in particular to a high-yielding avelamycin-producing strain and a breeding method thereof. Background technique [0002] Avilamycin (Avilamycin), also known as beptomycin, is an oligosaccharide antibiotic produced by the fermentation of Streptomyces viridoehrongenes. It was first developed by Eli Lilly and Company of the United States and named Sumisu (Maxus 100) is widely used for its high safety due to its unique structure, its easy degradation, almost no residue in the intestinal tract, low environmental pollution, low toxicity, and no cross-resistance. It is used as an additive in livestock and poultry feed, and there is no need for a withdrawal period. At present, avilamycin is widely used in major breeding countries in the world, such as the European Union, Japan, Brazil, etc., and was approved to be imported and used by the Ministry of Agriculture of my country in 2005 [Import...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N13/00C12N15/03C12R1/465
CPCC12N13/00C12N15/03C12N1/205C12R2001/465
Inventor 胡向东叶茂胡伟卿梁新乐
Owner 杭州皇冠农业生物工程技术研究中心有限公司
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