Abstract
A potent new lipopeptide antibiotic, A54145E(Asn3Asp9), was isolated from the fermentation broth of Streptomyces fradiae DA1489 engineered to delete genes encoding enzymes involved in hydroxylation of Asn3 and methoxylation of Asp9. The chemical structure predicted from the genetic changes in the biosynthetic pathway was determined by analyses of chemical transformations, D, L-amino acid quantitation by enantiomer labeling, tandem LC-MS/MS and 2D NMR techniques. These studies confirmed the primary amino acid sequence of A54145E(Asn3Asp9) predicted from the genetic engineering strategy, and also confirmed the structure and locations of three D-amino acids predicted from bioinformatic studies.
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Acknowledgements
We are grateful to C Mascio and J Silverman for MIC testing and H Cheng for initial LC-MS analyses. We also thank V Rajgarhia and C Gandhi for providing fermentation broth. We are also grateful to I Parsons and M Varoglu for their critical review of this manuscript. We acknowledge GJ Heffron of Harvard Medical School NMR facility for recording NMR spectra, WS Lane of Harvard Microchemistry and Proteomics analysis facility for standard amino acid analysis, and J Gerhardt of CAT GmbH, Tübingen, Germany for determination of optical purity of selected amino acids. This work was supported by Cubist Pharmaceuticals, Inc.
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Dedicated to the late Dr C Richard ‘Dick’/‘Hutch’ Hutchinson for his exceptional contributions to natural product biosynthesis, engineering, and drug discovery.
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Gu, JQ., Alexander, D., Rock, J. et al. Structural characterization of a lipopeptide antibiotic A54145E(Asn3Asp9) produced by a genetically engineered strain of Streptomyces fradiae. J Antibiot 64, 111–116 (2011). https://doi.org/10.1038/ja.2010.140
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DOI: https://doi.org/10.1038/ja.2010.140
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