Site-directed modification of the adenylation domain of the fusaricidin nonribosomal peptide synthetase for enhanced production of fusaricidin analogs

Jae Woo Han, Eun Young Kim, Jung Min Lee, Yun Sung Kim, Eunjung Bang, Beom Seok Kim

Research output: Contribution to journalArticle

22 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1327-1334
Number of pages8
JournalBiotechnology letters
Volume34
Issue number7
DOIs
Publication statusPublished - 2012 Jul

Keywords

  • Adenylation domain
  • Fusaricidin analogs
  • Non-ribosomal peptide synthetase
  • Paenibacillus polymyxa
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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