Department of Biological Science and Technology, University of Science and Technology Beijing, People's Republic of China.
Appl Biochem Biotechnol. 2010 Jan;160(2):393-400. doi: 10.1007/s12010-008-8324-y. Epub 2008 Aug 5.
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.
从具有高腈水合酶活性的红球菌 Rhodococcus rhodochrous tg1-A6 中，利用 PCR 扩增得到包含整个腈水合酶基因编码序列的 DNA 片段，并对其进行测序。腈水合酶基因的开放阅读框包含 1101 个碱基对，编码一个推测的 366 个氨基酸残基的多肽。将腈水合酶基因克隆到表达载体 pET-28a 中，并在大肠杆菌菌株 BL21(DE3)中表达。在重组菌中，检测到将各种腈转化为相应羧酸的腈水合酶的酶活性，在 9 小时时达到 24.5 U/ml。
