title: An FAD-dependent pyridine nucleotide-disulfide oxidoreductase is involved in disulfide bond formation in FK228 anticancer depsipeptide.
abstract: Disulfide bonds are rare in bacterial natural products, and the mechanism of disulfide bond formation in those products is unknown. Here we characterize a gene and its product critical for a disulfide bond formation in FK228 anticancer depsipeptide in Chromobacterium violaceum. Deletion of depH drastically reduced FK228 production, whereas complementation of the depH-deletion mutant with a copy of depH on a medium copy-number plasmid not only fully restored the FK228 production but also significantly increased the FK228 yield. Purified 6xHis-tagged DepH fusion protein in native form is a homodimer of 71.0 kDa, with each monomer containing one molecule of FAD. DepH efficiently converts an immediate FK228 precursor to FK228 in the presence of NADP(+). We conclude that DepH is an FAD-dependent pyridine nucleotide-disulfide oxidoreductase, specifically and efficiently catalyzing a disulfide bond formation in FK228.
authors: Department of Biological Sciences, University of Wisconsin-Milwaukee, PO Box 413, Milwaukee, WI 53201, USA.
authors: Cheng
authors: C
authors: Wang
authors: Shane R
authors: SR
authors: Wesener
authors: Hailong
authors: H
authors: Zhang
authors: Yi-Qiang
authors: YQ
authors: Cheng
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000378:metabolism*; D002861:Chromobacterium / Q000201:enzymology / Q000235:genetics; D047630:Depsipeptides / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny
chemical_list: D000903:Antibiotics, Antineoplastic; D047630:Depsipeptides; D004220:Disulfides; D050505:Mutant Proteins; D005182:Flavin-Adenine Dinucleotide; C087123:romidepsin; D009247:NADH, NADPH Oxidoreductases