Monday, July 27, 2009 - 11:00 AM
S16
Discovery and Engineering of Novel Enzymes and Pathways using DNA Assembler
Huimin Zhao, Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 S. Mathews Avenue, Urbana, IL 61801
Over the last decade, the complete genome sequences of nearly 1000 organisms have been determined. This provides a rich source for discovery of new natural products. However, only a tiny fraction of those putative natural product biosynthetic gene clusters have been characterized. Here we report the use of our newly developed DNA assembler method to discover, characterize, and engineer natural products. Unlike traditional cloning-based methods, this strategy synthesizes the entire expression plasmid containing the target gene cluster and the genetic elements needed for DNA maintenance and replication in S. cerevisiae, E. coli, and the target heterologous expression host in a single-step manner. As such, this strategy offers the greatest versatility and flexibility in manipulating the target gene cluster. As proof of concept, we used this approach to discover, characterize, and engineer several gene clusters including the fosfomycin gene cluster from Streptomyces fradiae, the spectinabilin gene cluster from Streptomyces orinoci, and multiple cryptic pathways from Streptomyces griseus. Along with these studies, we identified novel enzymes that can potentially be used as biocatalysts. One example is a new N-oxygenase that catalyzes oxidation of arylamines.
Web Page: www.chemeng.uiuc.edu/~zhaogrp/
See more of New tools for biocatalyst discovery (emphasis on tool development)
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See more of The Annual Meeting and Exhibition 2009 (July 26 - 30, 2009)
See more of Invited Oral Papers
See more of The Annual Meeting and Exhibition 2009 (July 26 - 30, 2009)