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J. Microbiol. Biotechnol. 2003; 13(5): 823-827

Published online October 28, 2003

Copyright © The Korean Society for Microbiology and Biotechnology.

Site-Directed Mutagenesis on Putative Macrolactone Ring Size Determinant in the Hybrid Pikromycin-Tylosin Polyketide Synthase

Jung Won-Seok , Eung-Soo Kim 1, Han-Young Kang 2, Cha-Yong Choi 3, David H. Sherman 4 and Yeo-Joon Yoon 5

Interdisciplinary Program of Biochemical Engineering and Biotechnology Seoul National University Seoul 151-742 Korea, 1School of Chemical Engineering and Biotechnology Inha University Incheon 402-751 Korea, 2Department of Chemistry College of Natural Science Chungbuk National University Cheongju Chungbuk 361-763 Korea, 3School of Chemical Engineering College of Engineering Seoul National University Seoul 151-742 Korea, 4Department of Microbiology and Biotechnology Institute University of Minnesota Minneapolis MN 55455 U.S.A., 5School of Chemical Engineering and Bioengineering University of Ulsan Ulsan 680-749 Korea

Abstract

Streptomyces venezuelae ATCC 15439 is notable in its ability to produce two distinct groups of macrolactones. It has been reported that the generation of two macrolactone structures results from alternative expression of pikromycin (Pik) polyketide synthase (PKS). It was previously reported that the hybrid pikromycin-tylosin PKS can also produce two different macrolactones but its mechanistic basis remains unclear. In order to address this question, a series of site-directed mutagenesis of tentative alternative ribosome binding site and translation start codons in tylGV were performed. The results suggest that macrolactone ring size is not determined by the alternative expression of TylGV but through other mechanism(s) involving direct interaction between the PikAIII and TE domain or skipping of the final chain elongation step. This provides new insight into the mechanism of macrolactone ring size determination in hybrid PKS as well as an opportunity to develop novel termination activities for combinatorial biosynthesis.

Keywords: Polyketide,polyketidesynthase,pikromycin,tylosin,domainskipping