Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 76, Issue 18, Pages 6295-6298Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.00029-10
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- South Korean government (MEST) [20090082333, 20090083328]
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Herewith we report the expression and screening of microbial enzymes without involving cloning procedures. Computationally predicted putative omega-transaminase (omega-TA) genes were PCR amplified from the bacterial colonies and expressed in a cell-free protein synthesis system for subsequent analysis of their enzymatic activity and substrate specificity. Through the cell-free expression analysis of the putative omega-TA genes, a number of enzyme-substrate pairs were identified in a matter of hours. We expect that the proposed strategy will provide a universal platform for bridging the information gap between nucleotide sequence and protein function to accelerate the discovery of novel enzymes.
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