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

Applied and Environmental Microbiology
Yong-Chan KwonDong-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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Citations

Feb 2, 2012·AMB Express·Jens RudatChristoph Syldatk
May 8, 2013·Biotechnology Advances·Christy CatherineDong-Myung Kim
Jan 13, 2015·Biotechnology Advances·Fabian Steffen-MunsbergUwe T Bornscheuer
Mar 3, 2015·Scientific Reports·Yong-Chan Kwon, Michael C Jewett
Apr 6, 2016·Synthetic and Systems Biotechnology·Jennifer A SchoborgMichael C Jewett
Jul 3, 2021·Life·Conary MeyerCheemeng Tan

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