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KMID : 0545120040140020356
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 2 p.356 ~ p.362
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Identification of a Cytochrome P450 Hydroxylase Gene Involved in Rifamycin Biosynthesis by Amycolatopsis mediterranei S699
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LEE, SANG-KIL
CHOI, CHA-YONG/AHN, JONG-SEOG/CHO, JAE-YONG/PARK, CHEON-SEOK/YOON, YEO JOON
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Abstract
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In analyzing the region of the Amycolatopsis mediterranei S699 chromosome responsible for the biosynthesis of the ansamycin antibiotic rifamycin, we identified a gene, designated orfO, which is located immediately upstream of the rifamycin polyketide synthase (PKS). OrfO encodes a protein, on the basis of sequence-comparative analysis, that is similar to several cytochrome P450 onooxygenases from different sources. The rifamycin producer, A. mediterranei, predominantly produces rifamycin B from its macrocyclic intermediate, proansamycin X, through dehydrogenation and hydroxylation steps. However, an A. mediterranei strain, deleted in orfO by gene replacement, no longer produced rifamycin B. Furthermore, a versatile replicative vector in A. mediterranei was constructed and rifamycin B production was restored in a complementation experiment of orfO using this novel vector. These consecutive results verified that the Orfo protein, which is a P450 hydroxylase, is required for the production of rifamycin B in A. mediterranei.
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