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

doi:10.1002/cbic.201500402

Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNA^UUA 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^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

33 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 language	English
Pages (from-to)	2498-2506
Number of pages	9
Journal	ChemBioChem
Volume	16
Issue number	17
Early online date	22 Oct 2015
DOIs	https://doi.org/10.1002/cbic.201500402
Publication status	Published - 23 Nov 2015

Keywords

bldA
fluorine
nucleocidin
Streptomyces calvus
sulfamate

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Zhu, X. M., Hackl, S., Thaker, M. N., Kalan, L., Weber, C., Urgast, D. S., Krupp, E. M., Brewer, A., Vanner, S., Szawiola, A., Yim, G., Feldmann, J., Bechthold, A., Wright, G. D., & Zechel, D. L. (2015). Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNA^UUA Molecule. ChemBioChem, 16(17), 2498-2506. https://doi.org/10.1002/cbic.201500402

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-tRNA^UUA Molecule', ChemBioChem, vol. 16, no. 17, pp. 2498-2506. https://doi.org/10.1002/cbic.201500402

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

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.",

keywords = "bldA, fluorine, nucleocidin, Streptomyces calvus, sulfamate",

author = "Zhu, {Xi Ming} and Stefanie Hackl and Thaker, {Maulik N.} and Lindsay Kalan and Claudia Weber and Urgast, {Dagmar S.} and Krupp, {Eva M.} and Alyssa Brewer and Stephanie Vanner and Anjuli Szawiola and Grace Yim and J{\"o}rg Feldmann and Andreas Bechthold and Wright, {Gerard D.} and Zechel, {David L.}",

note = "Funded by Natural Sciences and Engineering Research Council DFG EU Erasmus Fund Acknowledgements D.L.Z. and G.D.W. would like to thank the Natural Sciences and Engineering Research Council (NSERC) for financial support. We thank Dr. Bogdan Tokovenko, University of Saarbr{\"u}cken, for providing unpublished data on the transcription levels of genes involved in nucleocidin biosynthesis. This study was supported by the DFG (research training group 1976 grant to A.B.). C.W. thanks the EU Erasmus Fund for support.",

year = "2015",

month = nov,

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doi = "10.1002/cbic.201500402",

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AU - Zhu, Xi Ming

AU - Hackl, Stefanie

AU - Thaker, Maulik N.

AU - Kalan, Lindsay

AU - Weber, Claudia

AU - Urgast, Dagmar S.

AU - Krupp, Eva M.

AU - Brewer, Alyssa

AU - Vanner, Stephanie

AU - Szawiola, Anjuli

AU - Yim, Grace

AU - Feldmann, Jörg

AU - Bechthold, Andreas

AU - Wright, Gerard D.

AU - Zechel, David L.

N1 - Funded by Natural Sciences and Engineering Research Council DFG EU Erasmus Fund Acknowledgements D.L.Z. and G.D.W. would like to thank the Natural Sciences and Engineering Research Council (NSERC) for financial support. We thank Dr. Bogdan Tokovenko, University of Saarbrücken, for providing unpublished data on the transcription levels of genes involved in nucleocidin biosynthesis. This study was supported by the DFG (research training group 1976 grant to A.B.). C.W. thanks the EU Erasmus Fund for support.

PY - 2015/11/23

Y1 - 2015/11/23

N2 - 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.

AB - 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.

KW - bldA

KW - fluorine

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ER -

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

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