Asymmetrically simultaneous synthesis of L‐homophenylalanine and N6‐protected‐2‐oxo‐6‐amino‐hexanoic acid by engineered Escherichia coli aspartate aminotransferase |
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Authors: | Hsueh‐Hsia Lo Wei‐De Lin Shih‐Kuang Hsu Wen‐Hwei Hsu |
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Institution: | 1. Dept. of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung 406, Taiwan;2. Institute of Molecular Biology, National Chung‐Hsing University, Taichung 402, Taiwan;3. Dept. of Medical Research, China Medical University Hospital, Taichung 404, Taiwan;4. Dept. of Dental Laboratory Technology, Central Taiwan University of Science and Technology, Taichung 406, Taiwan |
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Abstract: | L ‐Homophenylalanine (L ‐HPA) and N6‐protected‐2‐oxo‐6‐amino‐hexanoic acid (N6‐protected‐OAHA) can be used as building blocks for the manufacture of angiotensin‐converting enzyme inhibitors. To synthesize L ‐HPA and N6‐protected‐OAHA simultaneously from 2‐oxo‐4‐phenylbutanoic acid (OPBA) and N6‐protected‐L ‐lysine, several variants of Escherichia coli aspartate aminotransferase (AAT) were developed by site‐directed mutagenesis and their catalytic activities were investigated. Three kinds of N6‐protected‐L ‐lysine were tested as potential amino donors for the bioconversion process. AAT variants of R292E/L18H and R292E/L18T exhibited specific activities of 0.70±0.01 U/mg protein and 0.67±0.02 U/mg protein to 2‐amino‐6‐tert‐butoxycarbonylamino‐hexanoic acid (BOC‐lysine) and 2‐amino‐6‐(2,2,2‐trifluoro‐acetylamino)‐hexanoic acid, respectively. E. coli cells expressing R292E/L18H variant were able to convert OPBA and BOC‐lysine to L ‐HPA and 2‐oxo‐6‐tert‐butoxycarbonylamino‐hexanoic acid (BOC‐OAHA) with 96.2% yield in 8 h. This is the first report demonstrating a process for the simultaneous production of two useful building blocks, L ‐HPA and BOC‐OAHA. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 |
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Keywords: | aspartate aminotransferase L‐homophenylanine bioconversion 2‐oxo‐6‐tertbutoxycarbonylamino‐hexanoic acid |
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