Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil |
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| Authors: | Yang Lu Ying Yu Rui Zhou Wei Sun Chunyan Dai Pei Wan Lanying Zhang Dongyun Hao Hejun Ren |
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| Affiliation: | (1) Key Laboratory of Water Resources and Environment of the Ministry of Education, College of Environment and Resources, Jilin University, 2519 Jiefang Road, Changchun, 130023, People’s Republic of China;(2) Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, 2699 Qianjin Street, Changchun, 130012, People’s Republic of China;(3) Biotechnology Research Centre, Jilin Academy of Agricultural Sciences (JAAS), 1363 Caiyu Street, Changchun, 130033, People’s Republic of China; |
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| Abstract: | 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 μM for 2,4-dichlorophenol and 6 μM for NADPH. The optimal temperature and pH of the enzyme were 25°C and 7.5, respectively. Activity of the purified TfdB-JLU was slightly enhanced by Ca2+, Mn2+, Co2+, and Fe2+, and completely inhibited by Cu2+, Hg2+, and Zn2+. This study is the first report to identify a novel TfdB from a metagenome. |
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