Cloning-independent expression and analysis of omega-transaminases by use of a cell-free protein synthesis system

利用无细胞蛋白合成系统进行ω-转氨酶的非克隆依赖性表达和分析

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作者:Yong-Chan Kwon,Kyung-Ho Lee, Ho-Cheol Kim, Kyuboem Han, Joo-Hyun Seo, Byung-Gee Kim, Dong-Myung Kim

Abstract

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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