Employing the over-expressed highly organic solvent tolerant alcohol dehydrogenase ADH-'A' from Rhodococcus ruber DSM 44541, versatile building blocks, which were not accessible by the wild type catalyst, were obtained in > 99% e.e.; furthermore, employing d8-2-propanol as deuterium source, stereoselective biocatalytic deuterium transfer was made feasible to furnish enantiopure deuterium labeled sec-alcohols on a preparative scale employing a single enzyme.

Biocatalytic hydrogen-transfer reduction of a-chloro-ketones furnished non-racemic chlorohydrins by employing either Rhodococcus ruber as lyophilized cell catalyst or an alcohol dehydrogenase preparation from Pseudomonas fluorescens DSM 50106 (PF-ADH). For all substrates investigated, Rhodococcus ruber gave strictly the "Prelog" product, whereas PF-ADH showed scattered stereopreference. One possibility for a follow-up reaction of halohydrins is the ring closure to the corresponding epoxide.A novel "one pot-one step strategy" was employed to obtain the enantiopure epoxide from the a-chloro-ketone in a cascade like fashion at pH > 12 involving biocatalytic hydrogen transfer reduction and in situ chemo-catalyzed ring closure.

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Contents General S2 General protocol for biocatalytic deuterium transfer, analytical scale S2 General protocol for biocatalytic deuterium transfer, preparative scale S3 Determination of the apparent kinetic isotope effect S3 Substrates and reference material S4 Sequencing of ADH-'A' and construction of the vector pET-22b+-ADH-'A' S7 Testing of various hosts S9 Optimized batch procedure for preparation of lyophilized cells of E. coli Tuner TM (DE3)/pET22b+-ADH-'A' S10 Analytics S11 Determination of absolute configuration S14 Optical Rotations S15 NMR spectra S16 References S27