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Cloning-Independent Expression and Analysis of ω-Transaminases by Use of a Cell-Free Protein Synthesis System

机译：使用无细胞蛋白质合成系统的ω-转氨酶的非克隆表达和分析

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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.
机译：因此，我们报道了不涉及克隆程序的微生物酶的表达和筛选。从细菌菌落中PCR扩增出计算预测的推定的ω-转氨酶（ω-TA）基因，并在无细胞蛋白质合成系统中表达，以便随后对其酶活性和底物特异性进行分析。通过对假定的ω-TA基因的无细胞表达分析，可以在数小时内鉴定出许多酶-底物对。我们希望所提出的策略将为弥合核苷酸序列和蛋白质功能之间的信息鸿沟提供一个通用平台，以加快新型酶的发现。

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