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Applied and Environmental Microbiology, August 2005, p. 4713-4720, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4713-4720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Engineering of Phenylacetaldehyde Reductase for Efficient Substrate Conversion in Concentrated 2-Propanol

Yoshihide Makino,* Kousuke Inoue, Tohru Dairi, and Nobuya Itoh

Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, Kosugi, Toyama 939-0398, Japan

Received 16 September 2004/ Accepted 13 March 2005

Phenylacetaldehyde reductase (PAR) is suitable for the conversion of various aryl ketones and 2-alkanones to corresponding chiral alcohols. 2-Propanol acts as a substrate solvent and hydrogen donor of coupled cofactor regeneration during the conversion of substrates catalyzed by PAR. To improve the conversion efficiency in high concentrations of substrate and 2-propanol, selection of a PAR mutant library and the subsequent rearrangement of mutations were attempted. With only a single selection round and following the manual combination of advantageous mutations, PAR was successfully adapted for the conversion of high concentrations of substrate with concentrated 2-propanol. This method will be widely applicable for the engineering of enzymes potentially valuable for industry.

* Corresponding author. Mailing address: Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, Kosugi, Toyama 939-0398, Japan. Phone: 81-766-56-7500, ext. 563. Fax: 81-766-56-2498. E-mail: makino{at}pu-toyama.ac.jp.

Applied and Environmental Microbiology, August 2005, p. 4713-4720, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4713-4720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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