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.
Müller
André
A
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
Hauer
Bernhard
B
Rosche
Bettina
B
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