Abstract
Identification of secondary metabolites produced by cryptic gene in bacteria may be difficult, but in the case of nonribosomal peptide (NRP)-type secondary metabolites, this study can be facilitated by bioinformatic analysis of the biosynthetic gene cluster and tandem mass spectrometry analysis. To illustrate this concept, we used mass spectrometry-guided bioinformatic analysis of genomic sequences to identify an NRP-type secondary metabolite from Streptomyces peucetius ATCC 27952. Five putative NRPS biosynthetic gene clusters were identified in the S. peucetius genome by DNA sequence analysis. Of these, the sp970 gene cluster encoded a complete NRPS domain structure, viz., C-A-T-C-A-T-E-C-A-T-C-A-T-C domains. Tandem mass spectrometry revealed that the functional siderophore peptide produced by this cluster had a molecular weight of 644.4 Da. Further analysis demonstrated that the siderophore peptide has a cyclic structure and an amino acid composition of AchfOrn–Arg–hOrn–hfOrn. The discovery of functional cryptic genes by analysis of the secretome, especially of NRP-type secondary metabolites, using mass spectrometry together with genome mining may contribute significantly to the development of pharmaceuticals such as hybrid antibiotics.
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Acknowledgments
This work was supported by the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Education, Science and Technology (2011-0031960) and Basic Science Research Program (2009-0071169) and Priority Research Centers Program (20110031388) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology.
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Park, HM., Kim, BG., Chang, D. et al. Genome-based cryptic gene discovery and functional identification of NRPS siderophore peptide in Streptomyces peucetius . Appl Microbiol Biotechnol 97, 1213–1222 (2013). https://doi.org/10.1007/s00253-012-4268-9
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DOI: https://doi.org/10.1007/s00253-012-4268-9