title: Site-directed modification of the adenylation domain of the fusaricidin nonribosomal peptide synthetase for enhanced production of fusaricidin analogs.
abstract: Fusaricidins produced by Paenibacillus polymyxa DBB1709 are lipopeptide antibiotics active against fungi and Gram-positive bacteria. The cyclic hexapeptide structures of fusaricidins are synthesized by fusaricidin synthetase, a non-ribosomal peptide synthetase. The adenylation domain of the third module (FusA-A3) can recruit L: -Tyr, L: -Val, L: -Ile, L: -allo-Ile, or L: -Phe, which diversifies the fusaricidin structures. Since the L: -Phe-incorporated fusaricidin analog (LI-F07) exhibits more potent antimicrobial activity than other analogs, we modified a specificity-conferring sequence in the substrate binding pocket of FusA-A3 to direct the enhanced production of LI-F07. Base on comparison to the adenylation domain of gramicidin S synthetase 1 and tyrocidine synthetase 1, both of which mainly activate L: -Phe, six mutant strains with altered FusA-A3 were generated using site-directed mutagenesis. M3 (I239W, I299V), M5 (I299V, G322A, V330I), and M6 (S239W, I299V, G322A, V330I) mutants produced significantly more LI-F07 than the wild-type strain.
authors.affiliation: Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Jae Woo
authors.initials: JW
authors.lastname: Han
authors.forename: Eun Young
authors.initials: EY
authors.lastname: Kim
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Lee
authors.forename: Yun Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Eunjung
authors.initials: E
authors.lastname: Bang
authors.forename: Beom Seok
authors.initials: BS
authors.lastname: Kim
mesh_terms: D019943:Amino Acid Substitution; D047630:Depsipeptides / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D056507:Paenibacillus / Q000201:enzymology*; D010453:Peptide Synthases / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary
chemical_list: D047630:Depsipeptides; D050505:Mutant Proteins; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase