Abstract
We describe an efficient method for producing both enantiomers of chiral alcohols by asymmetric hydrogen-transfer bioreduction of ketones in a 2-propanol (IPA)–water medium with E. coli biocatalysts expressing phenylacetaldehyde reductase (PAR: wild-type and mutant enzymes) from Rhodococcus sp. ST-10 and alcohol dehydrogenase from Leifsonia sp. S749 (LSADH). We also describe the detailed properties of mutant PARs, Sar268, and HAR1, which were engineered to have high activity and productivity in media composed of polar organic solvent and water, and the construction of three-dimensional structure of PAR by homology modeling. The K m and V max values for some substrates and the substrate specificity of mutant PARs were quite different from those of wild-type PAR. The results well explained the increased productivity of engineered PARs in IPA–water medium.
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Acknowledgments
This work was supported by a Grant-in-Aid for Scientific Research (B) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, and by a grant from the Japan Foundation for Applied Enzymology.
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An erratum to this article can be found at http://dx.doi.org/10.1007/s00253-012-3910-x.
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Itoh, N., Isotani, K., Nakamura, M. et al. Efficient synthesis of optically pure alcohols by asymmetric hydrogen-transfer biocatalysis: application of engineered enzymes in a 2-propanol–water medium. Appl Microbiol Biotechnol 93, 1075–1085 (2012). https://doi.org/10.1007/s00253-011-3447-4
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DOI: https://doi.org/10.1007/s00253-011-3447-4