Isolation of a novel gene encoding a 3,5,6-trichloro-2-pyridinol degrading enzyme from a cow rumen metagenomic library

• Published in: Biodegradation (Dordrecht) 2010-07, Vol.21 (4), p.565-573
• Main Authors: Math, Renukaradhya K, Asraful Islam, Shah Md, Cho, Kye Man, Hong, Sun Joo, Kim, Jong Min, Yun, Myoung Geun, Cho, Ji Joong, Heo, Jae Young, Lee, Young Han, Kim, Hoon, Yun, Han Dae
• Format: Article
• Language: English
• Subjects: 3,5,6-trichloro-2-pyridinol; Amidohydrolases - chemistry; Amidohydrolases - genetics; Amidohydrolases - metabolism; Amino Acid Sequence; Amino acids; Analysis; Animals; Aquatic Pollution; ATP; Bacteria - chemistry; Bacteria - enzymology; Bacteria - genetics; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Base Sequence; Biochemistry; Biodegradation; Biodegradation of pollutants; Biodegradation, Environmental; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cattle; Chlorpyrifos; Cloning, Molecular; E coli; Environment and pollution; Enzyme inhibitors; Enzymes; Escherichia coli; Fundamental and applied biological sciences. Psychology; Genes; Genomics; Geochemistry; High performance liquid chromatography; Industrial applications and implications. Economical aspects; Insecticides; Insecticides - metabolism; Libraries; Life Sciences; Liquid chromatography; Metabolites; Metagenomics; Microbiology; Molecular Sequence Data; Original Paper; Pesticides; Proteins; Pyridones - metabolism; Rumen; Rumen - chemistry; Rumen - microbiology; Rumen metagenome; Soil Science & Conservation; TCP degrading enzyme; Terrestrial Pollution; Transmission Control Protocol/Internet Protocol (Computer network protocol); Waste Management/Waste Technology; Waste Water Technology; Water Management; Water Pollution Control
• ISSN: 0923-9820; 1572-9729
• DOI: 10.1007/s10532-009-9324-5

3,5,6-trichloro-2-pyridinol (TCP) is a major metabolite of the insecticide chlorpyrifos and is hazardous to human and animal health. A gene encoding a TCP degrading enzyme was cloned from a metagenomic library prepared from cow rumen. The gene (tcp3A) is 2.5 kb in length, encoding a protein (Tcp3A) of 599 amino acid residues. Tcp3A has a potential signal sequence, as well as a putative ATP/GTP binding site, and a likely amidation site. The molecular weight of the enzyme is 62 kDa by SDS-PAGE. Comparison of Tcp3A with the NCBI database using BLASTP revealed homology to amidohydrolase proteins. Recombinant Escherichia coli harboring the tcp3A gene could utilize TCP as the sole source of carbon. TLC and HPLC revealed that TCP was degraded by recombinant E. coli harboring tcp3A. This is the first report of a gene encoding a TCP degrading enzyme.