Expression and characterization of a novel halohydrin dehalogenase from Tistrella mobilis KA081020-065 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 659-669, 2015.
Artículo
en Chino
| WPRIM
| ID: wpr-240611
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.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Rhodospirillaceae
/
Proteínas Bacterianas
/
Proteínas Recombinantes
/
Química
/
Clonación Molecular
/
Ácido 3-Hidroxibutírico
/
Escherichia coli
/
Genética
/
Hidrolasas
/
Hidroxibutiratos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2015
Tipo del documento:
Artículo
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