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ACS Chem. Biol.,
2 (6),
419–425
10.1021/cb700033s
Web Release Date: May 25, 2007
Copyright © 2007 American Chemical Society
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High-Throughput, Microarray-Based Synthesis of Natural Product Analogues via in Vitro Metabolic Pathway Construction
Seok Joon Kwon†, Moo-Yeal Lee‡, Bosung Ku†, David H. Sherman§, and Jonathan S. Dordick†,*
†Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180, ‡Solidus Biosciences, Inc., 1223 Peoples Avenue, Troy, New York 12180, §Life Sciences Institute, Department of Medicinal Chemistry, Chemical Biology Program, University of Michigan–Ann Arbor, 210 Washtenaw Avenue, Ann Arbor, Michigan 48109
Received for review February 14, 2007 and accepted April 26, 2007
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*Corresponding author, dordick#rpi.edu
The generation of biological diversity by engineering the biosynthetic gene assembly of metabolic pathway enzymes has led to a wide range of “unnatural” variants of natural products. However, current biosynthetic techniques do not allow the rapid manipulation of pathway components and are often fundamentally limited by the compatibility of new pathways, their gene expression, and the resulting biosynthetic products and pathway intermediates with cell growth and function. To overcome these limitations, we have developed an entirely in vitro approach to synthesize analogues of natural products in high throughput. Using several type III polyketide synthases (PKS) together with oxidative post-PKS tailoring enzymes, we performed 192 individual and multienzymatic reactions on a single glass microarray. Subsequent array-based screening with a human tyrosine kinase led to the identification of three compounds that acted as modest inhibitors in the low-micromolar range. This approach, therefore, enables the rapid construction of analogues of natural products as potential pharmaceutical lead compounds.
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