A kind of high-yield teicoplanin actinomycetes mutagenization strain and its application

Abstract

The present invention uses Actinoplanes teichomycoticus FIM‑68 (Actinoplanes teichomycoticus) as the starting strain, adopts the atmospheric pressure and room temperature plasma mutagenesis technology and screens the mutagenized strain obtained, and the mutagenized strain is Actinoplanes teichomycoticus FIM‑68‑66 (Actinoplanes teichomycoticus), which can ferment high-yield teicoplanin, greatly improves the yield of teicoplanin, and can be applied to industrial fermentation production; and the strain FIM‑68‑66 has good stability and continuous After three generations, the titer of substituted coplanin was basically stable and maintained at the same high level, so it could be used as a production strain for further research and development.

Description

【Technical field】 [0001] The invention belongs to the field of microorganisms, and in particular relates to a high-yield teicoplanin actinomycete mutagenesis strain and application thereof. 【Background technique】 [0002] Teicoplanin (teicoplanin) is a newly developed glycopeptide antibiotic against drug-resistant bacteria. Compared with vancomycin, an internationally recognized antibiotic against drug-resistant bacteria, teicoplanin has a similar antibacterial effect. activity, the same mechanism of action, similar or better clinical efficacy, and lower toxicity, especially lower nephrotoxicity. One of the active drugs. [0003] In the existing production methods, microbial fermentation is generally used to produce teicoplanin, and the crude product of teicoplanin obtained by fermentation is purified. Teicoplanin in the fermentation broth includes TAl~TA2, wherein TA2 is the main active ingredient of teicoplanin, mainly composed of 5 compounds with very similar chemical s...

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Problems solved by technology

[0004] At present, the breeding techniques for the improvement of teicoplanin strains mainly adopt breeding methods such as ultraviolet mutagenesis in combination with different mutagens, and protoplast fusion technology; for example: Cheong et al. isolated 1 strain mutant, called A. teichomyceticus BNG23, in order to further increase its production capacity, it was treated with ultraviolet light and obtained strain A. teichomyceticus BNG2315, the production capacity of this strain has reached 60 times that of the ATCC31121 strain, and it has been preserved in Korea. Species Preservation Center, No. KCCM-10601; Jin Zhihua et al. carried out ultraviolet mutagenesis to the starting bacterium A. teichomyceticus987-5-74 (preserved by Sichuan Antibiotics Culture Preservation Center), and used valine hydroxamic acid as a screening agent to obtain A resistant mutant strain A.teichomyceticus98-1-227, the total fermentation titer reached 2091μg / mL; The mutated spore suspension was coated on the separation medium containing 0.25% sodium acetate, and the sodium acetate-resistant mutant strain was screened, and the fermentation unit of the 04-10-3 strain obtained was 55% higher than that of the starting strain XYU-28; It can be seen from the above that ultraviolet mutagenesis combined with different mutagens has achieved certain effects. However, when these mutagens are used for a long time to treat the same strain, due to the non-directionality of mutations and the accumulation of mutations, while obtaining high yield, the strains have improved viability, Sporulation and other aspects will be significantly weakened, and multiple mutagenesis often leads to a high negative mutation rate and resistance saturation, so that there is insufficient room for strain improvement

Xu Bo et al. used genome shuffling technology to obtain 3 high-yield strains through mycelium preparation, protoplast fusion, and regeneration methods, among which the yield of SIIA05-03-136 strain (3016 μg / mL); although molecular biology The directional construction of functional cell lines based on genome rearrangement provides broad prospects, but it is extremely difficult to achieve the goal of transformation by culturing these functional cells; and due to the complexity and multi-node nature of the metabolic network in microorganisms, genetic manipulation The metabolic flow of the later strains is often not shifted in the direction of the expected design, especially in complex systems that produce secondary metabolites such as antibiotics, it is very difficult to obtain excellent industrial production strains

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