Abstract
Ramoplanins produced by Actinoplanes are new structural class of lipopeptide and are currently in phase III clinical trials for the prevention of vancomycin-resistant enterococcal infections. The depsipeptide structures of ramoplanins are synthesized by non-ribosomal peptide synthetases (NRPS). Romo-orf17, a stand-alone NRPS, is responsible for the recruitment of Thr into the linear NRPS pathways for which the corresponding adenylation domain is absent. Here, systematical gene inactivation and complementation have been carried out in a Actinoplanes sp. using homologous recombination and site-specific integration methods. A hybrid gene coding for the N-terminal region of the stand-alone NRPS and the A-PCP domains of a heterologous NRPS restored production of ramoplanins. The results elucidate the unusual N-terminal region which is essential for the biosynthesis of ramoplanins.
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Acknowledgments
This work was supported in part by grants from the National Natural Science Foundation of China (81072557, 81172962 and 30801449), the Science and Technology Commission of Shanghai Municipality (11QB1406300) and the Ministry of Science and Technology of China (Grant No. 2009ZX09301-007).
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Pan, HX., Li, JA., Shao, L. et al. Genetic manipulation revealing an unusual N-terminal region in a stand-alone non-ribosomal peptide synthetase involved in the biosynthesis of ramoplanins. Biotechnol Lett 35, 107–114 (2013). https://doi.org/10.1007/s10529-012-1056-7
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DOI: https://doi.org/10.1007/s10529-012-1056-7