期刊
TETRAHEDRON
卷 68, 期 37, 页码 7586-7590出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.tet.2012.06.058
关键词
BCAT; ilvE; L-Glutamate oxidase; Pichia pastoris; Escherichia coli
资金
- Federal Ministry of Economy, Family and Youth (BMWFJ)
- Federal Ministry of Traffic, Innovation and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol
- ZIT-Technology Agency of the City of Vienna through the COMET-Funding Program
Aminotransferases (ATs) are crucial enzymes for prokaryote and eukaryote metabolism, and have attracted industrial attention for large scale synthesis of amino acids and chiral amines. A high throughput screening procedure for branched chain aminotransferases (BCATs) was developed to identify optimal expression on a large number of expression clones. Escherichia coli BCAT, encoded by the ilvE gene, was expressed in E. coli and Pichia pastoris. This simple colorimetric assay procedure allowed the identification of optimal clones for ilvE expression and enabled the testing of its activity in cell lysates or on whole cell catalysis. (C) 2012 Elsevier Ltd. All rights reserved.
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