Abstract
A biosynthetic gene cluster for the 24-membered macrolactam antibiotic incednine was identified from the producer strain, Streptomyces sp. ML694-90F3. Among the putative incednine biosynthetic enzymes, a novel pyridoxal 5′-phosphate (PLP)-dependent β-glutamate-β-decarboxylase, IdnL3, was functionally characterized in vitro by demonstrating its (S)-3-aminobutyrate-forming activity with β-glutamate in the presence of PLP. Because (S)-3-aminobutyrate is known for the direct precursor of incednine, this enzyme supplies the unique β-amino acid starter unit. The identified gene cluster encodes five characteristic β-amino acid carrying enzymes, consisting of a pathway-specific ATP-dependent ligase, a discrete acyl carrier protein (ACP), β-aminoacyl-ACP β-amino group-protecting ATP-dependent ligase, dipeptidyl-ACP:PKS-loading ACP dipeptidyltransferase and a terminal amino acid peptidase, which are completely conserved in β-amino acid-containing macrolactam biosynthetic gene clusters. Overall, a plausible biosynthetic pathway for incednine was proposed.
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Acknowledgements
We thank Drs Sawa, Igarashi, Takahashi and Akamatsu at the Microbial Chemistry Research Center for providing us with the incednine producer Streptomyces sp. ML694-90F3, and Profs M Kobayashi (The University of Tsukuba, Japan) and K Arakawa (Hiroshima University, Japan) for providing plasmid pHSA81. This work was supported in part by Grants-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Sports, Science and Technology, the Nagase Science and Technology Foundation, and the Takeda Science Foundation.
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Takaishi, M., Kudo, F. & Eguchi, T. Identification of the incednine biosynthetic gene cluster: characterization of novel β-glutamate-β-decarboxylase IdnL3. J Antibiot 66, 691–699 (2013). https://doi.org/10.1038/ja.2013.76
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DOI: https://doi.org/10.1038/ja.2013.76
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