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Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil

文献类型： 外文期刊

作者： Lu, Yang 2 ; Yu, Ying 2 ; Zhou, Rui 1 ; Sun, Wei 1 ; Dai, Chunyan 2 ; Wan, Pei 2 ; Zhang, Lanying 1 ; Hao, Dongyun 2 ; Ren, 1 ;

作者机构： 1.Jilin Univ, Key Lab Water Resources & Environm, Minist Educ, Coll Environm & Resources, Changchun 130023, Peoples R China

2.Jilin Univ, Key Lab Mol Enzymol & Engn, Minist Educ, Changchun 130012, Peoples R China

3.JAAS, Biotechnol Res Ctr, Changchun 130033, Peoples R China

关键词： 2,4-Dichlorophenol hydroxylase;Herbicide degradation;Metagenomics;Polychlorobiphenyl

期刊名称：BIOTECHNOLOGY LETTERS （ 影响因子：2.461； 五年影响因子：2.457 ）

ISSN： 0141-5492

年卷期： 2011 年 33 卷 6 期

页码：

收录情况： SCI

摘要： A novel 2,4-dichlorophenol hydroxylase (TfdB, EC 1.14.13.20) gene, designated as tfdB-JLU, was identified from a metagenome constructed from polychlorinated biphenyl-contaminated soil by functional screening and heterologously expressed in Escherichia coli. The deduced amino acid sequence of tfdB-JLU exhibited less than 48% homology with other known TfdBs. The enzyme exhibited a wider substrate spectrum than the previously reported TfdBs and higher relative activity towards ortho-substituted dichlorophenols, 2-chlorophenol, and 3-chlorophenol than towards 2,4-dichlorophenol, the preferred substrate of other known TfdBs. The enzyme had a K(m) of 5 mu M for 2,4-dichlorophenol and 6 mu M for NADPH. The optimal temperature and pH of the enzyme were 25 degrees C and 7.5, respectively. Activity of the purified TfdB-JLU was slightly enhanced by Ca(2+), Mn(2+), Co(2+), and Fe(2+), and completely inhibited by Cu(2+), Hg(2+), and Zn(2+). This study is the first report to identify a novel TfdB from a metagenome.

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