Efficient expression in E. coli of an enantioselective nitrile hydratase from Rhodococcus erythropolis.
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 alpha- and beta-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 FeSO(4 )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.
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Liya
L
Song
Hong-Jie
HJ
Yuan
Lee
L
Coffey
John
J
Doran
Mei-Xiang
MX
Wang
Shijun
S
Qian
Catherine
C
O'Reilly
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics; D013237:Stereoisomerism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006836:Hydro-Lyases; C048831:nitrile hydratase