Isolation of putative polyene-producing actinomycetes strains via PCR-based genome screening for polyene-specific hydroxylase genes |
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| Institution: | 1. Department of Biotechnology, Inha University, Incheon 402-751, Republic of Korea;2. SongWon Envichem Inc., Kangnam-gu, Seoul 135-010, Republic of Korea;3. Hanson Biotech Co. Ltd., 201 IACRI, Han Nam University, Daeduk-gu, Daejeon 306-791, Republic of Korea;1. Chinese Center for Chirality, College of Pharmacy, Hebei University, Baoding 071002, China;2. The State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, CAS, Kunming 650204, China;3. Institut Pasteur, Unité de Chimie et Biocatalyse;4. CNRS, UMR 3523, 75724 Paris cedex 15, France;5. CNRS, UMR 5048, Centre de Biochimie Structurale, F34090 Montpellier, France;6. INSERM, U1054, F34090 Montpellier, France;1. Laboratory of Natural Products Chemistry, Department of Chemistry, Faculty of Science, University of Dschang, P.O. Box 67, Dschang, Cameroon;2. Institute of Natural Medicine, University of Toyama, 2630-Sugitani, Toyama 930-0194, Japan;3. Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho, Tokushima 770-8514, Japan |
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| Abstract: | Polyene antibiotics, which include nystatin, pimaricin, amphotericin and candicidin, include a family of very promising antifungal polyketide compounds that are typically produced by soil actinomycetes. The presence of similar cytochrome P450 hydroxylase (CYP) genes in the biosynthetic gene clusters for these polyenes have been previously reported. Using this polyene, more than 200 independently isolated actinomycetes strains were screened by CYP-specific PCR. Four strains were isolated based on the presence of the expected size of the PCR-amplified DNA fragment in the chromosome. The nucleotide sequencing of the PCR-amplified DNA fragments showed that each of the four actinomycetes strains contained a highly homologous polyene-specific CYP gene. Each of the culture extracts from these four strains showed a typical polyene-like high-pressure liquid chromatography (HPLC) chromatogram profile, and strong antifungal activity against Candida albicans. This suggests that the polyene-specific PCR-guided genome screening approach is an efficient method for isolating potentially valuable polyene-producing actinomycetes. |
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