|
|||||
|
|
Efficient expression in <Emphasis Type="Italic">E. coli</Emphasis> of an enantioselective nitrile hydratase from <Emphasis Type="Italic">Rhodococcus erythropolis</Emphasis> |
|
| Authors: | Liya Song Hong-Jie Yuan Lee Coffey John Doran Mei-Xiang Wang Shijun Qian Catherine O’Reilly |
| Institution: | (1) Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100080, China;(2) Graduate School of the Chinese Academy of Sciences, Beijing, 100039, China;(3) Department of Chemical and Life Sciences, Waterford Institute of Technology, Cork Road, Waterford, Ireland;(4) Laboratory of Chemical Biology, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100080, China |
| Abstract: | The genes encoding an enantioselective nitrile hydratase (NHase) from Rhodococcus erythropolis AJ270 have been cloned and an active NHase has been produced in Escherichia coli. Maximal activity was found when the genes encoding the α- and β-subunits were transcribed as one unit and the gene encoding the P44k activator protein as a separate ORF on a single replicon. Addition of n-butyric acid and FeSO4 could improve NHase activity. Coexpression of the GroEL-GroES chaperone proteins increased activity in the absence of P44k protein but had no effect in the presence of P44k. The recombinant enzyme was highly enantioselective in the synthesis of S-(+)-3-benzoyloxy- 4-cyanobutyramide from the prochiral substrate 3-benzoyloxyglutaronitrile. |
| Keywords: | Activator protein Enantioselectivity GroES-GroEL Nitrile hydratase Rhodococcus erythropolis |
| 本文献已被 PubMed SpringerLink 等数据库收录！ | |