Cloning-Independent Expression and Analysis of ω-Transaminases by Use of a Cell-Free Protein Synthesis System
2010 ◽
Vol 76
(18)
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pp. 6295-6298
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Keyword(s):
A Cell
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ABSTRACT Herewith we report the expression and screening of microbial enzymes without involving cloning procedures. Computationally predicted putative ω-transaminase (ω-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 ω-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.
1999 ◽
Vol 88
(3)
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pp. 345-347
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2008 ◽
Vol 101
(2)
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pp. 422-427
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2014 ◽
Vol 19
(3)
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pp. 426-432
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2015 ◽
Vol 20
(3)
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pp. 417-422
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1994 ◽
Vol 78
(2)
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pp. 130-133
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2009 ◽
Vol 102
(2)
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pp. 577-582
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