Asymmetric alkene reduction by yeast old yellow enzymes and by a novel Zymomonas mobilis reductase.
The genes encoding yeast old yellow enzymes (OYE 1, 2, and 3) and NAD(P)H-dependent 2-cyclohexen-1-one reductase from Zymomonas mobilis (NCR) were expressed separately in Escherichia coli. All four recombinant strains reduced the carbon double bond in alpha,beta-unsaturated alkenals and alkenones, however rates and enantio-specificities differed. Which of the two possible enantiomers was predominantly formed, was not only dependent on the choice of enzyme but also on the substrate: In addition to a dependency on methylation in alpha- or beta-position, the data of this study illustrate that firstly the E- or Z-configuration (cis- or trans-) of the carbon double-bond and secondly the remainder of the substrate molecule play roles in determining enantio-specificity. Based on the currently accepted mechanism of flavin mediated anti-hydrogenation of the carbon double bond, the data in this study may be explained by a flipped orientation of some of the substrates in the active center of OYE.
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
André
A
Müller
Bernhard
B
Hauer
Bettina
B
Rosche
D000475:Alkenes / Q000378:metabolism*; D001711:Biotransformation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007536:Isomerism; D009252:NADPH Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D012440:Saccharomyces / Q000235:genetics / Q000378:metabolism*; D017355:Zymomonas / Q000201:enzymology* / Q000235:genetics
D000475:Alkenes; D011994:Recombinant Proteins; D010088:Oxidoreductases; D009252:NADPH Dehydrogenase