A novel amidohydrolase (DmhA) from Sphingomonas sp that can hydrolyze the organophosphorus pesticide dimethoate to dimethoate carboxylic acid and methylamine
A novel amidohydrolase (DmhA) from Sphingomonas sp that can hydrolyze the organophosphorus pesticide dimethoate to dimethoate carboxylic acid and methylamine
复制标题
来自鞘氨醇单胞菌的新型酰胺水解酶 (DmhA)。
DOI：
10.1007/s10529-015-2027-6
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发表时间：
2016-04-01
影响因子：
2.7
通讯作者：
He, Jian
中科院分区：
文献类型：
--
作者：
Chen, Qing;Chen, Kai;He, Jian
Objectives:To characterize a novel dimethoate amidohydrolase from Sphingomonas sp. DC-6.Results:A gene, dmhA, encoding the dimethoate amidohydrolase responsible for transforming dimethoate to dimethoate carboxylic acid and methylamine, was cloned from Sphingomonas sp. DC-6. Sequence analysis and molecular modeling indicate that DmhA shares 31-57 % amino acid sequence identities with other functionally confirmed amidohydrolase. DmhA was expressed in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography. The purified DmhA could hydrolyze 4-acetaminophenol, dimethoate and propanil. DmhA activity was optimal at 30 °C and pH 7.5. Hg(2+), Zn(2+), Cu(2+), Cd(2+), Tween 80, Triton X-100 or SDS strongly inhibited its activity. The K m and k cat values of DmhA for dimethoate are 0.02 mM and 1.2 s(-1), respectively.Conclusions:DmhA was confirmed to be a novel dimethoate amidohydrolase which could eliminate the toxicity of dimethoate, providing a novel gene resource for the development of pesticide-degrading enzyme preparation and mechanistic study of dimethoate hydrolysis.
