Combined gene cluster engineering and precursor feeding to improve gougerotin production in Streptomyces graminearus
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Gougerotin is a peptidyl nucleoside antibiotic produced by Streptomyces graminearus. It is a specific inhibitor of protein synthesis and exhibits a broad spectrum of biological activities. Generation of an overproducing strain is crucial for the scale-up production of gougerotin. In this study, the natural and engineered gougerotin gene clusters were reassembled into an integrative plasmid by λ-red-mediated recombination technology combined with classic cloning methods. The resulting plasmids pGOU and pGOUe were introduced into S. graminearus to obtain recombinant strains Sgr-GOU and Sgr-GOUe, respectively. Compared with the wild-type strain, Sgr-GOU led to a maximum 1.3-fold increase in gougerotin production, while Sgr-GOUe resulted in a maximum 2.1-fold increase in gougerotin production. To further increase the yield of gougerotin, the effect of different precursors on its production was investigated. All precursors, including cytosine, serine, and glycine, had stimulatory effect on gougerotin production. The maximum gougerotin yield was achieved with Sgr-GOUe in the presence of glycine, and it was approximately 2.5-fold higher than that of the wild-type strain. The strategies used in this study can be extended to other Streptomyces for improving production of industrial important antibiotics.
KeywordsEngineering Precursor Gougerotin Gene cluster Streptomyces graminearus
This work was supported by grants from the National Natural Science Foundation of China (grant nos. 31171202 and 31270110) and the Ministry of Science and Technology of China (grant nos. 2012CB721103 and 2013CB734001). We would like to thank Dr. Bertolt Gust (University of Tübingen, Tübingen, Germany) for providing the PCR targeting system.
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