cis-Chlorobenzene dihydrodiol dehydrogenase (TcbB) from Pseudomonas sp strain P51, expressed in Escherichia coli DH5 alpha(pTCB149), catalyzes enantioselective dehydrogenase reactions

cis-Chlorobenzene dihydrodiol dehydrogenase (CDD) from Pseudomonas sp. stra in P51, cloned into Escherichia coli DH5 alpha(pTCB149) was able to oxidize cis-dihydrodihydroxy derivatives (cis-dihydrodiols) of dihydronaphthalene, indene, and four para-substituted toluenes to the corresponding catechols. During the incubation of a nonracemic mixture of cis-1,2-indandiol. only t he (+)-cis-(1R,2S) enantiomer was oxidized; the (-)-cis-(S,2R) enantiomer r emained unchanged. CDD oxidized both enantiomers of cis-1,2-dihydroxy-1,2,3 ,4-tetrahydronaphthalene, but oxidation of the (+)-cis-(1S,2R) enantiomer w as delayed until the (-)-cis-(1R,2S) enantiomer was completely depleted. Wh en incubated with nonracemic mixtures of para-substituted cis-toluene dihyd rodiols, CDD always oxidized the major enantiomer at a higher rate than the minor enantiomer. When incubated with racemic 1-indanol, CDD enantioselect ively transformed the (+)-(1S) enantiomer to 1-indanone. This stereoselecti ve transformation shows that CDD also acted as an alcohol dehydrogenase. Ad ditionally, CDD was able to oxidize (+)-cis-(1R,2S)-dihydroxy-1,2-dihydrona phthalene, (+)-cis-monochlorobiphenyl dihydrodiols, and (+)-cis-toluene dih ydrodiol to the corresponding catechols.