Expression and characterization of a novel halohydrin dehalogenase from Tistrella mobilis KA081020-065.
- Author:
Lei WANG
;
Jing YUAN
;
Peiyuan YAO
;
Lihua CHENG
;
Meixian XIE
;
Rongrong JIA
;
Huijin FENG
;
Min WANG
;
Qiaqing WU
;
Dunming ZHU
- Publication Type:Journal Article
- MeSH:
3-Hydroxybutyric Acid;
chemistry;
Bacterial Proteins;
genetics;
metabolism;
Cloning, Molecular;
Escherichia coli;
Hydrolases;
genetics;
metabolism;
Hydroxybutyrates;
chemistry;
Recombinant Proteins;
genetics;
metabolism;
Rhodospirillaceae;
enzymology;
genetics
- From:
Chinese Journal of Biotechnology
2015;31(5):659-669
- CountryChina
- Language:Chinese
-
Abstract:
Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
