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Chembiochem. 2015 Nov;16(17):2498-506. doi: 10.1002/cbic.201500402. Epub 2015 Oct 22.

Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNA(UUA) Molecule.

Author information

1
Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada.
2
Department of Pharmaceutical Biology and Biotechnology, Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104, Freiburg, Germany.
3
Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1200 Main Street West, Hamilton, Ontario, L8N 3Z5, Canada.
4
Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB24 3UE, UK.
5
Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada. dlzechel@chem.queensu.ca.

Abstract

Nucleocidin is one of the very few natural products known to contain fluorine. Mysteriously, the nucleocidin producer Streptomyces calvus ATCC 13382 has not been observed to synthesize the compound since its discovery in 1956. Here, we report that complementation of S. calvus ATCC 13382 with a functional bldA-encoded Leu-tRNA(UUA) molecule restores the production of nucleocidin. Nucleocidin was detected in culture extracts by (19) F NMR spectroscopy, HPLC-ESI-MS, and HPLC-continuum source molecular absorption spectroscopy for fluorine-specific detection. The molecule was purified from a large-scale culture and definitively characterized by NMR spectroscopy and high-resolution MS. The nucleocidin biosynthetic gene cluster was identified by the presence of genes encoding the 5'-O-sulfamate moiety and confirmed by gene disruption. Two of the genes within the nucleocidin biosynthetic gene cluster contain TTA codons, thus explaining the dependence on bldA and resolving a 60-year-old mystery.

KEYWORDS:

Streptomyces calvus; bldA; fluorine; nucleocidin; sulfamate

PMID:
26374477
DOI:
10.1002/cbic.201500402
[Indexed for MEDLINE]

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