title: Use of a phosphonate methyltransferase in the identification of the fosfazinomycin biosynthetic gene cluster.
abstract: Natural product discovery has been boosted by genome mining approaches, but compound purification is often still challenging. We report an enzymatic strategy for "stable isotope labeling of phosphonates in extract" (SILPE) that facilitates their purification. We used the phosphonate methyltransferase DhpI involved in dehydrophos biosynthesis to methylate a variety of phosphonate natural products in crude spent medium with a mixture of labeled and unlabeled S-adenosyl methionine. Mass-guided fractionation then allowed straightforward purification. We illustrate its utility by purifying a phosphonate that led to the identification of the fosfazinomycin biosynthetic gene cluster. This unusual natural product contains a hydrazide linker between a carboxylic acid and a phosphonic acid. Bioinformatic analysis of the gene cluster provides insights into how such a structure might be assembled.
authors.affiliation: Institute for Genomic Biology, University of Illinois at Urbana-Champaign (USA).
authors.forename: Jiangtao
authors.initials: J
authors.lastname: Gao
authors.forename: Kou-San
authors.initials: KS
authors.lastname: Ju
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Yu
authors.forename: Juan E
authors.initials: JE
authors.lastname: Velásquez
authors.forename: Subha
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Jaeheon
authors.initials: J
authors.lastname: Lee
authors.forename: Changming
authors.initials: C
authors.lastname: Zhao
authors.forename: Bradley S
authors.initials: BS
authors.lastname: Evans
authors.forename: James R
authors.initials: JR
authors.lastname: Doroghazi
authors.forename: William W
authors.initials: WW
authors.lastname: Metcalf
authors.forename: Wilfred A
authors.initials: WA
authors.lastname: van der Donk
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D001688:Biological Products / Q000737:chemistry / Q000378:metabolism*; D019295:Computational Biology; D004271:DNA, Fungal / Q000235:genetics; D006834:Hydrazines / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D007553:Isotope Labeling; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D016366:Open Reading Frames / Q000235:genetics; D063065:Organophosphonates / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D012436:S-Adenosylmethionine / Q000737:chemistry; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001688:Biological Products; D004271:DNA, Fungal; D006834:Hydrazines; D063065:Organophosphonates; D009943:Organophosphorus Compounds; C000591602:fosfazinomycin; C029424:hydrazine; D012436:S-Adenosylmethionine; D008780:Methyltransferases
keywords: antibiotics; biosynthesis; hydrazine; natural products