Purification and characterization of nitrile hydratase of mutant 4D of Rhodococcus rhodochrous PA-34

被引：6

作者：

Pratush, Amit ^{[1
]}

Seth, Amit ^{[1
]}

Bhalla, Tek Chand ^{[1
]}

机构：

[1] Himachal Pradesh Univ, Dept Biotechnol, Shimla 171005, Himachal Prades, India

来源：

3 BIOTECH | 2013年 / 3卷 / 02期

关键词：

Characterization; Mutant; Purification; Rhodococcus rhodochrous PA-34;

D O I：

10.1007/s13205-012-0081-5

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Nitrile hydratase (NHase; E.C. 4.2.1.84) has been purified and characterized using ammonium sulfate precipitation, ion exchange chromatography and gel filtration chromatography from the mutant 4D of Rhodococcus rhodochrous PA-34. The SDS-PAGE and MALDI-TOF analysis of the purified enzyme revealed that it is dimmer consisting of alpha- and beta-subunits with a molecular mass of 25 and 30 kDa, respectively. The K-m and V-max values were 102 mM and 350.8 mu mol/min/mg using 3-cyanopyridine as substrate. The purified NHase was stable in higher concentration of potassium ions and in acidic pH 5.5 as compared to NHase of the wild R. rhodochrous PA-34. The analysis of the N-terminal amino acid sequence of this enzyme revealed that this enzyme has 90 % homology with the high molecular weight nitrile hydratase of R. rhodochrous J1.

引用

页码：165 / 171

页数：7

共 26 条

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