Identification of the nonribosomal peptide synthetase gene responsible for bassianolide synthesis in wood-decaying fungus Xylaria sp. BCC1067
| Autor: | Jiraporn Jirakkakul, Anuwat Tachaleat, Morakot Tanticharoen, Somchai Pongpattanakitshote, Supapon Cheevadhanarak, Porntip Paungmoung, Namol Vorapreeda, Cheeranun Klomnara, Juntira Punya |
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| Rok vydání: | 2008 |
| Předmět: |
Magnetic Resonance Spectroscopy
Sequence analysis Mutant Molecular Sequence Data Antitubercular Agents Mutagenesis (molecular biology technique) Antineoplastic Agents Microbial Sensitivity Tests Biology Microbiology Peptides Cyclic Polymerase Chain Reaction Open Reading Frames Nonribosomal peptide Cell Line Tumor Chlorocebus aethiops Animals Humans Peptide Synthases DNA Fungal Gene Chromatography High Pressure Liquid DNA Primers Genetics chemistry.chemical_classification Molecular Structure Sequence Homology Amino Acid Xylariales Fungal genetics Sequence Analysis DNA Wood Major facilitator superfamily Anti-Bacterial Agents Protein Structure Tertiary Open reading frame Mutagenesis Insertional Biochemistry chemistry Gene Deletion |
| Zdroj: | Microbiology (Reading, England). 154(Pt 4) |
| ISSN: | 1350-0872 |
| Popis: | Intensive study of gene diversity of bioactive compounds in a wood-rot fungus, Xylaria sp. BCC1067, has made it possible to identify polyketides and nonribosomal peptides (NRPs) unaccounted for by conventional chemical screening methods. Here we report the complete nonribosomal peptide synthetase (NRPS) gene responsible for the biosynthesis of an NRP, bassianolide, using a genetic approach. Isolation of the bassianolide biosynthetic gene, nrpsxy, was achieved using degenerate primers specific to the adenylation domain of NRPS. The complete ORF of nrpsxy is 10.6 kb in length. Based on comparisons with other known NRPSs, the domain arrangement of NRPSXY is most likely to be C-A-T-C-A-M-T-T-C-R. The other ORF found upstream of nrpsxy, designated efxy, is 1.8 kb in length and shows high similarity to members of the major facilitator superfamily of transporters. Functional analysis of the nrpsxy gene was conducted by gene disruption, and the missing metabolite in the mutant was identified. Chemical analysis revealed the structure of the metabolite to be a cyclooctadepsipeptide, bassianolide, which has been found in other fungi. A bioassay of bassianolide revealed a wide range of biological activities other than insecticidal uses, which have been previously reported, thus making bassianolide an interesting candidate for future structural modification. This study is the first evidence for a gene involved in the biosynthesis of bassianolide. |
| Databáze: | OpenAIRE |
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