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Method for acquiring erythromycin streptomycete high-yield strain

A technology of high-yield strains and erythromycin, applied in the fields of gene mutagenesis and gene recombination, can solve problems such as cell chromosome changes and increase the appearance of high-yield strains, and achieve the effects of improving metabolic capacity, broadening genetic germplasm resources, and good economic benefits

Inactive Publication Date: 2013-06-19
SHANDONG FANGMING PHARMACEUTICAL CO LTD
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Problems solved by technology

Moreover, protoplast fusion can often cause the elimination of cell plasmids, which can lead to changes in cell chromosomes, or changes in secondary metabolic pathways, thereby increasing the chance of high-yielding strains

Method used

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Examples

Experimental program
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Embodiment Construction

[0029] The present invention will be further described below by implementation example, and following example is illustrative, not limiting, can not limit protection scope of the present invention with following example.

[0030] one. Ultraviolet and fast neutron mutagenesis to obtain high-yield strains Ⅰ and Ⅱ

[0031] 1. Screening of starting strains

[0032] Use an inoculation shovel to take an appropriate amount of sand containing Streptomyces erythromycin from the sand tube, put it into a triangular flask filled with glass beads, vibrate vigorously to break up the spores, and spread them on the plate respectively according to the gradient after gradient dilution, 34± Cultivate at 1°C, humidity 50% for 9-11 days. Pick 10 single colonies from the plate and transfer them to the slant, and transfer them into the shake flask after maturity to determine their titer (Table 1). The strain SE7 with high titer and relatively stable was selected as the starting strain (Table 2). ...

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Abstract

The invention relates to a method for acquiring an erythromycin streptomycete high-yield strain. The method is characterized in that erythromycin streptomycete is respectively mutated by using ultraviolet rays and fast neutron, such that high-yield strains I and II are obtained; with a protoplast fusion technology, the two high-yield strains are fused, such that a novel high-yield recombinant is obtained. A shaking flask titer of the recombinant is 6513u / ml, which is improved by 14.5% than an original strain, and is respectively improved by 8.9% and 5.7% than the mutant strains I and II. After times of passages, the titer is stable, and a small-scale fermentation titer can be higher than 11000u / ml. With the method, strain metabolism is effectively improved, and good economic benefit is obtained. The method is an ideal microbial strain breeding method.

Description

technical field [0001] The present invention relates to the fields of gene mutagenesis and gene recombination, specifically manifested as ultraviolet mutagenesis and fast neutron mutagenesis technology, and protoplast fusion technology to obtain high-yield strains, which not only improves the economic benefits of enterprises, but also broadens the genetics of microorganisms. Germplasm resources enrich the content of the gene bank. Background technique [0002] At present, the fermentation level of erythromycin-producing bacteria is low, and the production cost is relatively high, which cannot meet the needs of today's market. Therefore, breeding high-yielding strains of erythromycin is a key problem to be solved in the production and development of erythromycin. At present, conventional physical and chemical factor mutation breeding is widely used in the breeding of erythromycin-producing bacteria. Mutation breeding can improve the production capacity of strains to a certa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00C12N1/21C12N15/03C12R1/465
Inventor 李瑾程传东何海燕张丽丽于新令王凤滩
Owner SHANDONG FANGMING PHARMACEUTICAL CO LTD