RRC ID 15809
著者 Wang H, Liu N, Xi L, Rong X, Ruan J, Huang Y.
タイトル Genetic screening strategy for rapid access to polyether ionophore producers and products in actinomycetes.
ジャーナル Appl Environ Microbiol
Abstract Polyether ionophores are a unique class of polyketides with broad-spectrum activity and outstanding potency for the control of drug-resistant bacteria and parasites, and they are produced exclusively by actinomycetes. A special epoxidase gene encoding a critical tailoring enzyme involved in the biosynthesis of these compounds has been found in all five of the complete gene clusters of polyether ionophores published so far. To detect potential producer strains of these antibiotics, a pair of degenerate primers was designed according to the conserved regions of the five known polyether epoxidases. A total of 44 putative polyether epoxidase gene-positive strains were obtained by the PCR-based screening of 1,068 actinomycetes isolated from eight different habitats and 236 reference strains encompassing eight major families of Actinomycetales. The isolates spanned a wide taxonomic diversity based on 16S rRNA gene analysis, and actinomycetes isolated from acidic soils seemed to be a promising source of polyether ionophores. Four genera were detected to contain putative polyether epoxidases, including Micromonospora, which has not previously been reported to produce polyether ionophores. The designed primers also detected putative epoxidase genes from diverse known producer strains that produce polyether ionophores unrelated to the five published gene clusters. Moreover, phylogenetic and chemical analyses showed a strong correlation between the sequence of polyether epoxidases and the structure of encoded polyethers. Thirteen positive isolates were proven to be polyether ionophore producers as expected, and two new analogues were found. These results demonstrate the feasibility of using this epoxidase gene screening strategy to aid the rapid identification of known products and the discovery of unknown polyethers in actinomycetes.
巻・号 77(10)
ページ 3433-42
公開日 2011-5-1
DOI 10.1128/AEM.02915-10
PII AEM.02915-10
PMID 21421776
PMC PMC3126441
MeSH Actinobacteria / genetics* Actinobacteria / metabolism* Biosynthetic Pathways / genetics Cluster Analysis DNA Primers / genetics DNA, Bacterial / chemistry DNA, Bacterial / genetics* DNA, Ribosomal / chemistry DNA, Ribosomal / genetics Ionophores / metabolism* Mass Screening / methods* Molecular Sequence Data Multigene Family Oxidoreductases / genetics Phylogeny Polymerase Chain Reaction / methods* Polymers / metabolism* RNA, Ribosomal, 16S / genetics Sequence Analysis, DNA
IF 4.016
引用数 13
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
一般微生物 JCM 4703 JCM 3308 JCM 3373 JCM 12949 JCM 7543 JCM 12951 JCM 9535 JCM 9312 JCM 9537 JCM 9536 JCM 9534 JCM 3246 JCM 3195 JCM 11711 JCM 3090 JCM 12343 JCM 11001 JCM 11002 JCM 6844 JCM 3313 JCM 9103 JCM 3378 JCM 3005 JCM 3092 JCM 3109 JCM 3145 JCM 3295 JCM 3309