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Efficient Biosynthesis of Ethyl ( R )‐3‐Hydroxyglutarate through a One‐Pot Bienzymatic Cascade of Halohydrin Dehalogenase and Nitrilase
Author(s) -
Yao Peiyuan,
Wang Lei,
Yuan Jing,
Cheng Lihua,
Jia Rongrong,
Xie Meixian,
Feng Jinhui,
Wang Min,
Wu Qiaqing,
Zhu Dunming
Publication year - 2015
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201500061
Subject(s) - nitrilase , chemistry , biotransformation , biosynthesis , escherichia coli , substrate (aquarium) , mutant , solvent , stereochemistry , enzyme , biochemistry , nuclear chemistry , organic chemistry , gene , biology , ecology
An effective one‐pot bienzymatic synthesis of ethyl ( R )‐3‐hydroxyglutarate (EHG) from ethyl ( S )‐4‐chloro‐3‐hydroxybutyrate (ECHB) was achieved by using recombinant Escherichia coli cells expressing separately or co‐expressing a mutant halohydrin dehalogenase gene from Agrobacterium radiobacter AD1 and a nitrilase gene from Arabidopsis thaliana . The activity of nitrilase was inhibited by high concentration of ECHB and NaCN. Consequently, the one‐pot one‐step process was implemented by fed‐batch of ECHB and NaCN with high accumulative product concentration (up to 0.9 mol L −1 ). The biotransformation of ECHB to EHG was successfully achieved at 1.2 mol L −1 substrate concentration by a one‐pot two‐step process. As such, this one‐pot bienzymatic transformation should be useful in synthesizing these important optical pure β‐hydroxycarboxylic acids.