Abstract
Nikkomycins and gougerotin are peptidyl nucleoside antibiotics with broad biological activities. The nikkomycin biosynthetic gene cluster comprises one pathway-specific regulatory gene (sanG) and 21 structural genes, whereas the gene cluster for gougerotin biosynthesis includes one putative regulatory gene, one major facilitator superfamily transporter gene, and 13 structural genes. In the present study, we introduced sanG driven by six different promoters into Streptomyces ansochromogenes TH322. Nikkomycin production was increased significantly with the highest increase in engineered strain harboring hrdB promoter-driven sanG. In the meantime, we replaced the native promoter of key structural genes in the gougerotin (gou) gene cluster with the hrdB promoters. The heterologous producer Streptomyces coelicolor M1146 harboring the modified gene cluster produced gougerotin up to 10-fold more than strains carrying the unmodified cluster. Therefore, genetic manipulations of genes involved in antibiotics biosynthesis with the constitutive hrdB promoter present a robust, easy-to-use system generally useful for the improvement of antibiotics production in Streptomyces.
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Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (grant nos. 31171202 and 31030003) and the Ministry of Science and Technology of China (grant nos. 2013CB734001 and 2012CB721103). We would like to thank Professor Mervyn Bibb (John Innes Centre, Norwich, UK) for providing S. coelicolor M1146. We also thank Dr. Bertolt Gust (University of Tübingen, Tübingen, Germany) for providing the PCR targeting system and Professor Keqian Yang (the Institute of Microbiology, Chinese Academy of Sciences, Beijing, China) for the gift of pIMEP.
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Deyao Du and Yu Zhu contributed equally to this work.
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Du, D., Zhu, Y., Wei, J. et al. Improvement of gougerotin and nikkomycin production by engineering their biosynthetic gene clusters. Appl Microbiol Biotechnol 97, 6383–6396 (2013). https://doi.org/10.1007/s00253-013-4836-7
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DOI: https://doi.org/10.1007/s00253-013-4836-7