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- Title
Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil.
- Authors
Yang Lu; Ying Yu; Rui Zhou; Wei Sun; Chunyan Dai; Pei Wan; Lanying Zhang; Dongyun Hao; Hejun Ren
- Abstract
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 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 Ca, Mn, Co, and Fe, and completely inhibited by Cu, Hg, and Zn. This study is the first report to identify a novel TfdB from a metagenome.
- Subjects
GENETIC engineering; DICHLOROPHENOXYACETIC acid; CLONING; CHOLESTEROL hydroxylase; POLYCHLORINATED biphenyls; ESCHERICHIA coli
- Publication
Biotechnology Letters, 2011, Vol 33, Issue 6, p1159
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-011-0549-0