Asymmetric reduction of 2-chloroacrylic acid to (S)-2-chloropropionic acid by a novel reductase from Burkholderia sp. WS
Graphical abstract
Introduction
Biologically active compounds, including pharmaceutical and agrochemical products, often contain stereogenic centers. In most cases, one of the enantiomers has the desired biological activity, whereas the other does not or is even hazardous. Thus, there exists a continuing demand for the development of technology to produce optically active compounds efficiently.
(S)-2-Chloropropionic acid (CPA) is used as a building block for the synthesis of aryloxyphenoxypropionic acid, the (R)-isomer of which has herbicidal activity. In the industrial synthesis of this herbicide, (S)-2-CPA is produced by resolution, in which the (R)-2-CPA component of a racemic 2-CPA mixture is selectively degraded using (R)-2-haloacid dehalogenase.1 Since the theoretical maximum yield of (S)-2-CPA from a racemic mixture of 2-CPA by this procedure is only 50%, it would be advantageous to establish an asymmetric synthesis procedure that could produce (S)-2-CPA directly. Herein, we have developed an enzymatic process in which (S)-2-CPA is produced by asymmetric reduction of the carbon–carbon double bond in 2-chloroacrylic acid (2-CAA).
Section snippets
Results and discussion
We have previously isolated bacterial strains that use 2-CAA as their sole carbon and energy source.2 One of these strains, formerly called Pseudomonas sp. WS, was used herein. We re-characterized this strain by 16S rRNA sequencing and found that the sequence had the highest similarity to those of bacteria belonging to the genus Burkholderia. Thus, we have renamed this strain Burkholderia sp. WS.
Burkholderia sp. WS exhibits 2-CPA dehalogenase activity when grown in media containing 2-CPA as the
Conclusion
In summary, we found that 2-CAA-grown Burkholderia sp. WS produces a novel enzyme that catalyzes the asymmetric reduction of 2-CAA to (S)-2-CPA. This enzyme required NADPH as a co-substrate. Since (S)-2-CPA is probably further metabolized by (S)-DEX, deletion of the (S)-DEX gene would be essential in order to accumulate (S)-2-CPA in Burkholderia sp. WS. Cloning of the gene encoding this novel enzyme, as well as the overproduction and further characterization of the gene product, are currently
Acknowledgements
This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas 13125203 (to N.E.), a Grant-in-Aid for Young Scientists (A) 14703021 (to T.K.) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the National Project on Protein Structural and Functional Analyses, and Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (21st Century COE on Kyoto University Alliance for Chemistry).
References (7)
- et al.
J. Biol. Chem.
(1995) - et al.
J. Biol. Chem.
(1997) - et al.
Angew. Chem., Int. Ed.
(2004)
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