title: Gene cloning, overexpression, and characterization of the nitrilase from Rhodococcus rhodochrous tg1-A6 in E. coli.
abstract: A DNA fragment containing the entire coding sequence of nitrilase gene was amplified from Rhodococcus rhodochrous tg1-A6 with high nitrilase activity using PCR and sequenced. The open reading frame of the nitrilase gene contains 1,101 base pairs, which encodes a putative polypeptide of 366 amino acid residues. The nitrilase gene was cloned into an expression vector pET-28a and expressed in an Escherichia coli strain BL21(DE3). The enzymatic activity of nitrilase, which converts various nitriles to the corresponding carboxylic acids, was detected to reach 24.5 U/ml at 9 h in the recombinant bacteria.
authors.affiliation: Department of Biological Science and Technology, University of Science and Technology Beijing, People's Republic of China. luohui99@yahoo.com.cn
authors.forename: Hui
authors.initials: H
authors.lastname: Luo
authors.forename: Lu
authors.initials: L
authors.lastname: Fan
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Chang
authors.forename: Jinwei
authors.initials: J
authors.lastname: Ma
authors.forename: Huimin
authors.initials: H
authors.lastname: Yu
authors.forename: Zhongyao
authors.initials: Z
authors.lastname: Shen
mesh_terms: D000619:Aminohydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D012240:Rhodococcus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D000619:Aminohydrolases; C021867:nitrilase