Expression of the Gene of Glycerol Dehydrogenase from Hansenula polymorpha Dl-1 in Escherichia coli for the Production of Chiral Compounds

For the production of chiral compounds, an Escherichia coli HB101 strain was transformed. A chiral column that could resolve the three 2,3-butanediol isomers and the two acetoin isomers was used to demonstrate the stereospecificity of the enzyme. The H. polymorpha DL-1 gene product, glycerol dehydrogenase, catalyses the NAD+-dependent oxidation of 2,3-butanediol to acetoin as well as the corresponding reverse reactions. The recombinant E. coli HB101 strain harbouring the expression plasmid produced (3R)-acetoin [99% yield, >99.9% enantiomeric excess (e.e.)] from 110 mM (2R, 3R)-2,3-butanediol and (3S)-acetoin (99% yield, >99.9% e.e.) from 110 mM meso-2,3-butanediol after 24 h of incubation, showing specificity towards the secondary alcohol in R-configuration. From a racemate of 110 mM 2,3-butanediol (the molar ratio of 17 (2R, 3R) : 15 (2S, 3S) : 78 (meso)), (2S, 3S)-2,3-butanediol (15 mM, 92% e.e.) was obtained in the resting-cell reaction without any additive to regenerate NAD+ from NADH.