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Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA ‐Specified Leu‐tRNA UUA Molecule
Author(s) -
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.
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500402
Subject(s) - gene cluster , biosynthesis , moiety , stereochemistry , chemistry , nuclear magnetic resonance spectroscopy , streptomyces , complementation , natural product , molecule , small molecule , reverse genetics , transfer rna , gene , biochemistry , biology , rna , organic chemistry , genetics , bacteria , mutant , phenotype
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.