Peric-Concha, Nataša and Borovicka, Branko and Long, Paul F. and Hranueli, Daslav and Waterman, Peter G. and Hunter, Iain S. (2005) Ablation of the OtcC gene encoding a post-polketide hydroxylase from the oxytetracyline biosynthetic pathway in streptomyces rimosus results in novel polyketides with altered chain length. Journal of Biological Chemistry, 280. pp. 37455-37460. ISSN 1083-351XFull text not available in this repository. (Request a copy from the Strathclyde author)
Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC “minimal” polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex.
|Keywords:||oxytetracycline, ablated gene, synthase complex, Pharmacy and materia medica, Biochemistry, Cell Biology, Molecular Biology|
|Subjects:||Medicine > Pharmacy and materia medica|
|Department:||Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences|
|Depositing user:||Pure Administrator|
|Date Deposited:||22 Feb 2012 16:20|
|Last modified:||03 Feb 2017 12:28|