LI Yang, TANG Lixia, ZHENG Kai, et al. Optimizing expression and purification of recombinant halohydrin dehalogenase from A.radiobacter AD1[J]. CIESC Journal, 2010, (12): 3213-3219.
LI Yang, TANG Lixia, ZHENG Kai, et al. Optimizing expression and purification of recombinant halohydrin dehalogenase from A.radiobacter AD1[J]. CIESC Journal, 2010, (12): 3213-3219.DOI:
Optically pure epoxides and β-substituted alcohols
constituting one of the most important classes of building blocks for the synthesis of chiral drugs
could be obtained via reactions catalyzed by halohydrin dehloagenases.Researches aimed at the improvement of over-expression and purification of recombinant halohydrin dehalogenases will greatly facilitate its application.The encoding gene was successfully cloned into pBAD expression vector.Under the developed optimized expression conditions
halohydrin dehalogenase could be expressed as a soluble format in E.coli using the two expression systems.The soluble targeted protein was about 35% of the total soluble proteins as judged by activity measurements and SDS-PAGE analysis.From 0.5 L cultures of each
34 mg and 41 mg of 90% pure halohydrin dehalogenase were obtained by using one-step Q-Sepharose anion exchange chromatography
respectively.Meanwhile
the constructed pBADHheC plasmid would set a solid base for improving the biocatalytic properties of the enzyme.