数字农科院2.0

Overexpression of wysR gene enhances wuyiencin production in Delta wysR3 mutant strain of Streptomyces albulus var. wuyiensis strain CK-15

文献类型： 外文期刊

作者： Wei, Q.;Aung, A.;Liu, B.;Ma, J.;Shi, L.;Zhang, K.;Ge, B.;Aung, A.

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关键词： biocontrol; Gibson assembly; overexpression; wuyiencin; wysR; Delta wysR3

期刊名称： JOURNAL OF APPLIED MICROBIOLOGY

ISSN： 1364-5072

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收录情况： SCI

摘要： Aim: The aim of the present work was to investigate the overexpression of thewysRgene inStreptomyces albulusvar. wuyiensis strain CK-15 based on the Delta wysR3 mutant strain including the effect on morphological development, wuyiencin production and antibacterial activity. At the same time, we report a new rapid method for producing genetically engineered strains for industrial production of wuyiencin. Methods and Results: We developed a method to create awysRoverexpression strain based on the Delta wysR3 mutant strain by direct transformation. In this method, the desired gene fragment to be overexpressed was amplified by polymerase chain reaction (PCR) using Phusion High Fidelity DNA polymerase and fused with the linearized pSETC integrative plasmid by Gibson assembly. The resulting recombinant plasmid was transformed into Delta wysR3 mutant strain by the intergeneric conjugation method. The plasmid was then integrated into the chromosome and the resulting apramycin-resistant overexpression strain was confirmed by PCR using the Apra-F and Apra-R primers. Finally, we successfully screened the genetically engineered strain with overexpression of wys Rgene in Delta wysR3 mutant. Conclusion: We can conclude that overexpression of wysRgene in Delta wysR3 mutant strain proved to be an effective strategy for significantly increasing wuyienc in production together with faster morphological development. Quantitative real-time RT-PCR analysis showed that wysR regulated wuyiencin biosynthesis by modulating other putative regulatory genes and bld,whi,chp,rdl and ram family genes are crucial for the morphological development. Significance and Impact of the Study: Overexpression of wysR gene in the Delta wysR3 mutant strain named OoWysR strain may increase the efficiency in the industrial fermentation processes for wuyiencin production. The mechanism by which wysR overexpression promotes rapid sporulation and a high yield of wuyiencin production is likely related to modulation of other putative regulatory genes.

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