L. Jones et al., PURIFICATION AND CHARACTERIZATION OF D-BETA-HYDROXYBUTYRATE DEHYDROGENASE EXPRESSED IN ESCHERICHIA-COLI, Biochemistry and cell biology, 71(7-8), 1993, pp. 406-410
D-beta-Hydroxybutyrate dehydrogenase (BDH), a lipid-requiring enzyme,
has been cloned into pUC18, expressed in Escherichia coli, and purifie
d to homogeneity. The apoenzyme, i.e., the enzyme devoid of phospholip
id, has no activity, but can be activated by phospholipid to a specifi
c activity of 129 mu mol/(min mg). The functional properties of the en
zyme expressed in E. coil were compared with the enzyme purified from
rat liver. The specific activities, kinetic parameters, and phospholip
id activation profiles were virtually identical. These results indicat
e that the expression of the enzyme in E. coli is a viable method for
producing active functional BDH and should allow for the production of
specifically altered BDH molecules.