An exceptionally DMSO-tolerant alcohol dehydrogenase for the stereoselective reduction of ketones.
A novel short-chain alcohol dehydrogenase from Paracoccus pantotrophus DSM 11072, which is applicable for hydrogen transfer, has been identified, cloned, and overexpressed in E. coli. The enzyme stereoselectively reduces several ketones in a sustainable substrate-coupled approach using 2-propanol (5% v/v) as hydrogen donor. The enzyme maintained its activity in organic co-solvents in biphasic as well as monophasic systems and was even active in micro-aqueous media (1% v/v aqueous buffer). In general, a higher conversion was observed at higher log P values of the solvent, however, DMSO, which exhibits the lowest log P value of all solvents investigated, was not only tolerated but led to a higher conversion and relative activity (110-210%). For example, the conversion after 24 h in 15% v/v DMSO was double that for the reaction performed in buffer. This tolerance to DMSO may be attributed to the ability of the wild-type strain to adapt and grow in media with high sulfur content.
Lavandera
Iván
I
Research Centre Applied Biocatalysis c/o Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria.
Kern
Alexander
A
Schaffenberger
Martina
M
Gross
Johannes
J
Glieder
Anton
A
de Wildeman
Stefaan
S
Kroutil
Wolfgang
W
D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D055162:Biocatalysis; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology; D007659:Ketones / Q000737:chemistry* / Q000378:metabolism*; D015394:Molecular Structure; D010084:Oxidation-Reduction; D012997:Solvents; D013237:Stereoisomerism
D007659:Ketones; D012997:Solvents; D000426:Alcohol Dehydrogenase; D004121:Dimethyl Sulfoxide