Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNAUUA Molecule

Xi Ming Zhu, Stefanie Hackl, Maulik N. Thaker, Lindsay Kalan, Claudia Weber, Dagmar S. Urgast, Eva M. Krupp, Alyssa Brewer, Stephanie Vanner, Anjuli Szawiola, Grace Yim, Jörg Feldmann, Andreas Bechthold, Gerard D. Wright, David L. Zechel

Research output: Contribution to journalArticle

13 Citations (Scopus)

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‐tRNAUUAmolecule restores the production of nucleocidin. Nucleocidin was detected in culture extracts by 19F 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.

Original languageEnglish
Pages (from-to)2498-2506
Number of pages9
JournalChemBioChem
Volume16
Issue number17
Early online date22 Oct 2015
DOIs
Publication statusPublished - 23 Nov 2015

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Biosynthesis
Streptomyces
Biological Products
Molecules
Genes
Multigene Family
Spectrum Analysis
Molecular spectroscopy
Gene encoding
Fluorine
Absorption spectroscopy
Codon
Nuclear magnetic resonance spectroscopy
nucleocidin
Magnetic Resonance Spectroscopy
Spectroscopy

Keywords

  • bldA
  • fluorine
  • nucleocidin
  • Streptomyces calvus
  • sulfamate

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNAUUA Molecule. / Zhu, Xi Ming; Hackl, Stefanie; Thaker, Maulik N.; Kalan, Lindsay; Weber, Claudia; Urgast, Dagmar S.; Krupp, Eva M.; Brewer, Alyssa; Vanner, Stephanie; Szawiola, Anjuli; Yim, Grace; Feldmann, Jörg; Bechthold, Andreas; Wright, Gerard D.; Zechel, David L.

In: ChemBioChem, Vol. 16, No. 17, 23.11.2015, p. 2498-2506.

Research output: Contribution to journalArticle

Zhu, XM, Hackl, S, Thaker, MN, Kalan, L, Weber, C, Urgast, DS, Krupp, EM, Brewer, A, Vanner, S, Szawiola, A, Yim, G, Feldmann, J, Bechthold, A, Wright, GD & Zechel, DL 2015, 'Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNAUUA Molecule' ChemBioChem, vol. 16, no. 17, pp. 2498-2506. https://doi.org/10.1002/cbic.201500402
Zhu, Xi Ming ; Hackl, Stefanie ; Thaker, Maulik N. ; Kalan, Lindsay ; Weber, Claudia ; Urgast, Dagmar S. ; Krupp, Eva M. ; Brewer, Alyssa ; Vanner, Stephanie ; Szawiola, Anjuli ; Yim, Grace ; Feldmann, Jörg ; Bechthold, Andreas ; Wright, Gerard D. ; Zechel, David L. / Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNAUUA Molecule. In: ChemBioChem. 2015 ; Vol. 16, No. 17. pp. 2498-2506.
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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‐tRNAUUAmolecule restores the production of nucleocidin. Nucleocidin was detected in culture extracts by 19F 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.",
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