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| A gene cluster for biosynthesis of the sesquiterpenoid antibiotic pentalenolactone in Streptomyces avermitilis |
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| Author(s): Tetzlaff CN, You Z, Cane DE, Takamatsu S, Omura S, Ikeda H |
| Source: BIOCHEMISTRY Volume: 45 Issue: 19 Pages: 6179-6186 Published: MAY 16 2006 |
| Times Cited: 15 References: 43 |
| Abstract: Streptomyces avermitilis, an industrial organism responsible for the production of the anthelminthic avermectins, harbors a 13.4 kb gene cluster containing 13 unidirectionally transcribed open reading frames corresponding to the apparent biosynthetic operon for the sesquiterpene antibiotic pentalenolactone. The advanced intermediate pentalenolactone F, along with the shunt metabolite pentalenic acid, could be isolated from cultures of S. avermitilis, thereby establishing that the pentalenolactone biosynthetic pathway is functional in S. avermitilis. Deletion of the entire 13.4 kb cluster from S. avermitilis abolished formation of pentalenolactone metabolites, while transfer of the intact cluster to the pentalenolactone nonproducer Streptomyces lividans 1326 resulted in production of pentalenic acid. Direct evidence for the biochemical function of the individual biosynthetic genes came from expression of the ptlA gene (SAV2998) in Escherichia coli. Assay of the resultant protein established that PtlA is a pentalenene synthase, catalyzing the cyclization of farnesyl diphosphate to pentalenene, the parent hydrocarbon of the pentalenolactone family of metabolites. The most upstream gene in the cluster, gap1 (SAV2990), was shown to correspond to the pentalenolactone resistance gene, based on expression in E. coli and demonstration that the resulting glyceraldehyde-3-phosphate dehydrogenase, the normal target of pentalenolactone, was insensitive to the antibiotic. Furthermore, a second GAPDH isozyme (gap2, SAV6296) has been expressed in E. coli and shown to be inactivated by pentalenolactone. |
| Document Type: Article |
| Language: English |
| Reprint Address: Cane, DE (reprint author), Brown Univ, Dept Chem, Box H, Providence, RI 02912 USA |
Addresses:
1. Brown Univ, Dept Chem, Providence, RI 02912 USA
2. Kitasato Univ, Kitasato Inst Life Sci, Sagamihara, Kanagawa 2288555 Japan
3. Kitasato Inst, Minato Ku, Tokyo 1088642, Japan |
| Publisher: AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USA |
| Subject Category: Biochemistry & Molecular Biology |
| IDS Number: 043DG |
| ISSN: 0006-2960 |
| DOI: 10.1021/bi060419n |
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