CLONING, NUCLEOTIDE-SEQUENCE AND EXPRESSION IN ESCHERICHIA-COLI OF 2 COBALT-CONTAINING NITRILE HYDRATASE GENES FROM RHODOCOCCUS-RHODOCHROUS J1

被引:105
作者
KOBAYASHI, M [1 ]
NISHIYAMA, M [1 ]
NAGASAWA, T [1 ]
HORINOUCHI, S [1 ]
BEPPU, T [1 ]
YAMADA, H [1 ]
机构
[1] UNIV TOKYO,FAC AGR,DEPT AGR CHEM,TOKYO 113,JAPAN
基金
日本学术振兴会;
关键词
NITRILE HYDRATASE GENE; CLONING; COBALT; (RHODOCOCCUS); (ESCHERICHIA-COLI);
D O I
10.1016/0167-4781(91)90208-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Rhodococcus rhodochrous J1 produces two kinds of cobalt-containing nitrile hydratases (NHases); one is a high molecular mass-NHase (H-NHase) and the other is a low molecular mass-NHase (L-NHase). Both NHases are composed of two subunits of different sizes (alpha and beta-subunits). The H- and L-NHase genes were cloned into Escherichia coli by a DNA-probing method using the NHase gene of Rhodococcus sp. N-774, a ferric ion-containing NHase producing strain, as the hybridization probe and their nucleotide sequences were determined. In each of the H- and L-NHase genes, the open reading frame for the beta-subunit was located just upstream of that for the alpha-subunit, which probably belongs to the same operon. The amino acid sequences of each subunit of the H- and L-NHases from R. rhodochrous J1 showed generally significant similarities to those from Rhodococcus sp. N-774, but the arrangement of the coding sequences for two subunits is reverse of the order found in the NHase gene of Rhodococcus sp. N-774. Each of the NHase genes was expressed in E. coli cells under the control of lac promoter, only when they were cultured in the medium supplemented with CoCl2.
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页码:23 / 33
页数:11
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