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Research Paper |
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain1
Author for correspondence: José A. Gil. Tel: +34 987 291503. Fax: +34 987 291479. e-mail: degjgs{at}unileon.es
A 205 kb DNA region from Streptomyces griseus IMRU 3570, including the candicidin biosynthetic gene cluster, was cloned and partially sequenced. Analysis of the sequenced DNA led to identification of genes encoding part of a modular polyketide synthase (PKS), genes for thioesterase, macrolactone ring modification, mycosamine biosynthesis and attachment to the macrolide ring, candicidin export and regulatory proteins. It represents the first extensive genetic characterization of an aromatic polyene macrolide antibiotic biosynthetic gene cluster. Of particular interest is the presence of the CanP1 loading domain (the first described as responsible for the activation of an aromatic starter unit) and the polypeptide CanP3 (carrying modules for the formation of five out of seven conjugated double bonds). Disruption of the pabAB gene that encodes the starter unit of candicidin abolished its production [which was restored when exogenous p-aminobenzoic acid (PABA) was supplied to the culture] and resulted in an enhanced production of another antifungal compound that is barely detected in the wild-type.
Keywords: aromatic polyene antibiotic, type I PKS, genetic disruption
Abbreviations: ACP, acyl carrier protein; AT, acyltransferase; DH, dehydratase; ER, enoylreductase; KR, ß-ketoreductase; KS, ß-ketoacyl ACP synthase; mAT, methylmalonate-specific AT; PABA, p-aminobenzoic acid; PKS, polyketide synthase; TE, thioesterase
The GenBank accession numbers for the sequences reported in this paper are AJ300302 and AJ300303.
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