Detection of nonribosomal peptide synthetase genes in Xylaria sp. BCC1067 and cloning of XyNRPSA.
Nonribosomal peptides, synthesized by nonribosomal peptide synthetases (NRPS), are an important group of diverse bioactive fungal metabolites. Xylaria sp. BCC1067, which is known to produce a variety of biologically active metabolites, was studied for gene encoding NRPS by two different PCR-based methods and seven different NRPS fragments were obtained. In addition, screening a genomic library with an amplified NRPS fragment as a probe identified a putative NRPS gene named XyNRPSA. The functionality of XyNRPSA for the production of a corresponding metabolite was probed by gene insertion inactivation. Comparing the disrupting metabolite profile with that of the wild type led to the identification of a speculated metabolite. The crude extract of Xylaria sp. BCC1067 also exhibits antifungal activity against the human pathogens Candida albicans and Trichophyton mentagrophytes. However, the evaluation of biological activity of the XyNRPSA product suggests that it is neither a compound with antifungal activity nor a siderophore. In the vicinity of XyNRPSA, a second gene (named XyPtB) was identified. Its localization and homology to orfB of the ergot alkaloid biosynthetic gene cluster suggests that XyPtB may be involved in XyNRPSA product biosynthesis.
Paungmoung
Porntip
P
Department of Biotechnology, School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand. porntip19@yahoo.com
Punya
Jantira
J
Pongpattanakitshote
Somchai
S
Jeamton
Wattana
W
Vichisoonthonkul
Taweerat
T
Bhumiratana
Sakarindr
S
Tanticharoen
Morakot
M
Linne
Uwe
U
Marahiel
Mohamed A
MA
Cheevadhanarak
Supapon
S
D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D000935:Antifungal Agents / Q000378:metabolism; D003001:Cloning, Molecular; D010453:Peptide Synthases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010456:Peptides, Cyclic / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D013098:Xylariales / Q000201:enzymology / Q000235:genetics*
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D010456:Peptides, Cyclic; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase