Identification of Genes Essential for Sulfamate and Fluorine Incorporation During Nucleocidin Biosynthesis

A. R.Ola Pasternak; Andreas Bechthold; David L. Zechel

doi:10.1002/cbic.202200140

Identification of Genes Essential for Sulfamate and Fluorine Incorporation During Nucleocidin Biosynthesis

A. R.Ola Pasternak
, Andreas Bechthold
, David L. Zechel

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

12 Scopus citations

Abstract

Nucleocidin is an adenosine derivative containing 4’-fluoro and 5’-O-sulfamoyl substituents. In this study, nucleocidin biosynthesis is examined in two newly discovered producers, Streptomyces virens B-24331 and Streptomyces aureorectus B-24301, which produce nucleocidin and related derivatives at titers 30-fold greater than S. calvus. This enabled the identification of two new O-acetylated nucleocidin derivatives, and a potential glycosyl-O-acetyltransferase. Disruption of nucJ, nucG, and nucI, within S. virens B-24331, specifying a radical SAM/Fe−S dependent enzyme, sulfatase, and arylsulfatase, respectively, led to loss of 5’-O-sulfamoyl biosynthesis, but not fluoronucleoside production. Disruption of nucN, nucK, and nucO specifying an amidinotransferase, and two sulfotransferases respectively, led to loss of fluoronucleoside production. Identification of S. virens B-24331 as a genetically tractable and high producing strain sets the stage for understanding nucleocidin biosynthesis and highlights the utility of using 16S-RNA sequences to identify alternative producers of valuable compounds in the absence of genome sequence data.

Original language	English
Article number	e202200140
Journal	ChemBioChem
Volume	23
Issue number	15
DOIs	https://doi.org/10.1002/cbic.202200140
State	Published - Aug 3 2022
Externally published	Yes

Funding

D.L.Z. and A.R.O.P are grateful to the Natural Sciences and Engineering Research Council (NSERC) of Canada for financial support (03695-2016). A.R.O.P. is additionally grateful to Mitacs for the Globalink Research Travel Award. Prof. Dr. Andriy Luzhetskyy (Saarbrücken University) is thanked for a generous gift of the plasmids pKC1132 and pUWL−H. We are also very grateful to Dr. Francoise Sauriol for help performing the NMR experiments. D.L.Z. and A.R.O.P are grateful to the Natural Sciences and Engineering Research Council (NSERC) of Canada for financial support (03695‐2016). A.R.O.P. is additionally grateful to Mitacs for the Globalink Research Travel Award. Prof. Dr. Andriy Luzhetskyy (Saarbrücken University) is thanked for a generous gift of the plasmids pKC1132 and pUWL−H. We are also very grateful to Dr. Francoise Sauriol for help performing the NMR experiments.

Keywords

biosynthesis
fluorine
gene disruption
nucleocidin
sulfamate

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10.1002/cbic.202200140

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title = "Identification of Genes Essential for Sulfamate and Fluorine Incorporation During Nucleocidin Biosynthesis",

abstract = "Nucleocidin is an adenosine derivative containing 4{\textquoteright}-fluoro and 5{\textquoteright}-O-sulfamoyl substituents. In this study, nucleocidin biosynthesis is examined in two newly discovered producers, Streptomyces virens B-24331 and Streptomyces aureorectus B-24301, which produce nucleocidin and related derivatives at titers 30-fold greater than S. calvus. This enabled the identification of two new O-acetylated nucleocidin derivatives, and a potential glycosyl-O-acetyltransferase. Disruption of nucJ, nucG, and nucI, within S. virens B-24331, specifying a radical SAM/Fe−S dependent enzyme, sulfatase, and arylsulfatase, respectively, led to loss of 5{\textquoteright}-O-sulfamoyl biosynthesis, but not fluoronucleoside production. Disruption of nucN, nucK, and nucO specifying an amidinotransferase, and two sulfotransferases respectively, led to loss of fluoronucleoside production. Identification of S. virens B-24331 as a genetically tractable and high producing strain sets the stage for understanding nucleocidin biosynthesis and highlights the utility of using 16S-RNA sequences to identify alternative producers of valuable compounds in the absence of genome sequence data.",

keywords = "biosynthesis, fluorine, gene disruption, nucleocidin, sulfamate",

author = "Pasternak, \{A. R.Ola\} and Andreas Bechthold and Zechel, \{David L.\}",

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year = "2022",

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AU - Pasternak, A. R.Ola

AU - Bechthold, Andreas

AU - Zechel, David L.

PY - 2022/8/3

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N2 - Nucleocidin is an adenosine derivative containing 4’-fluoro and 5’-O-sulfamoyl substituents. In this study, nucleocidin biosynthesis is examined in two newly discovered producers, Streptomyces virens B-24331 and Streptomyces aureorectus B-24301, which produce nucleocidin and related derivatives at titers 30-fold greater than S. calvus. This enabled the identification of two new O-acetylated nucleocidin derivatives, and a potential glycosyl-O-acetyltransferase. Disruption of nucJ, nucG, and nucI, within S. virens B-24331, specifying a radical SAM/Fe−S dependent enzyme, sulfatase, and arylsulfatase, respectively, led to loss of 5’-O-sulfamoyl biosynthesis, but not fluoronucleoside production. Disruption of nucN, nucK, and nucO specifying an amidinotransferase, and two sulfotransferases respectively, led to loss of fluoronucleoside production. Identification of S. virens B-24331 as a genetically tractable and high producing strain sets the stage for understanding nucleocidin biosynthesis and highlights the utility of using 16S-RNA sequences to identify alternative producers of valuable compounds in the absence of genome sequence data.

AB - Nucleocidin is an adenosine derivative containing 4’-fluoro and 5’-O-sulfamoyl substituents. In this study, nucleocidin biosynthesis is examined in two newly discovered producers, Streptomyces virens B-24331 and Streptomyces aureorectus B-24301, which produce nucleocidin and related derivatives at titers 30-fold greater than S. calvus. This enabled the identification of two new O-acetylated nucleocidin derivatives, and a potential glycosyl-O-acetyltransferase. Disruption of nucJ, nucG, and nucI, within S. virens B-24331, specifying a radical SAM/Fe−S dependent enzyme, sulfatase, and arylsulfatase, respectively, led to loss of 5’-O-sulfamoyl biosynthesis, but not fluoronucleoside production. Disruption of nucN, nucK, and nucO specifying an amidinotransferase, and two sulfotransferases respectively, led to loss of fluoronucleoside production. Identification of S. virens B-24331 as a genetically tractable and high producing strain sets the stage for understanding nucleocidin biosynthesis and highlights the utility of using 16S-RNA sequences to identify alternative producers of valuable compounds in the absence of genome sequence data.

KW - biosynthesis

KW - fluorine

KW - gene disruption

KW - nucleocidin

KW - sulfamate

UR - https://www.scopus.com/pages/publications/85130437929

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DO - 10.1002/cbic.202200140

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AN - SCOPUS:85130437929

SN - 1439-4227

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JO - ChemBioChem

JF - ChemBioChem

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ER -

Identification of Genes Essential for Sulfamate and Fluorine Incorporation During Nucleocidin Biosynthesis

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