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Li R,
Oliver RA,
Townsend CA,
( 2017 ) Identification and Characterization of the Sulfazecin Monobactam Biosynthetic Gene Cluster. PMID : 28017601 : DOI : 10.1016/j.chembiol.2016.11.010 PMC : PMC5286544 Abstract >>
The monobactams, exemplified by the natural product sulfazecin, are the only class of �]-lactam antibiotics not inactivated by metallo-�]-lactamases, which confer bacteria with extended-spectrum �]-lactam resistance. We screened a transposon mutagenesis library from Pseudomonas acidophila ATCC 31363 and isolated a sulfazecin-deficient mutant that revealed a gene cluster encoding two non-ribosomal peptide synthetases (NRPSs), a methyltransferase, a sulfotransferase, and a dioxygenase. Three modules and an aberrant C-terminal thioesterase (TE) domain are distributed across the two NRPSs. Biochemical examination of the adenylation (A) domains provided evidence that L-2,3-diaminopropionate, not L-serine as previously thought, is the direct source of the �]-lactam ring of sulfazecin. ATP/PPi exchange assay also revealed an unusual substrate selectivity shift of one A domain when expressed with or without the immediately upstream condensation domain. Gene inactivation analysis defined a cluster of 13 open reading frames sufficient for sulfazecin production, precursor synthesis, self-resistance, and regulation. The identification of a key intermediate supported a proposed NRPS-mediated mechanism of sulfazecin biosynthesis and �]-lactam ring formation distinct from the nocardicins, another NRPS-derived subclass of monocyclic �]-lactam. These findings will serve as the basis for further biosynthetic research and potential engineering of these important antibiotics.
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