Isolation of a novel gene encoding a 3,5,6-trichloro-2-pyridinol degrading enzyme from a cow rumen metagenomic library.
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.
Math
Renukaradhya K
RK
Division of Applied Life Science (BK21 Program), Gyeongsang National University, Chinju, 660-701, Republic of Korea.
Asraful Islam
Shah Md
SM
Cho
Kye Man
KM
Hong
Sun Joo
SJ
Kim
Jong Min
JM
Yun
Myoung Geun
MG
Cho
Ji Joong
JJ
Heo
Jae Young
JY
Lee
Young Han
YH
Kim
Hoon
H
Yun
Han Dae
HD
D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001673:Biodegradation, Environmental; D002417:Cattle; D003001:Cloning, Molecular; D007306:Insecticides / Q000378:metabolism; D056186:Metagenomics; D008969:Molecular Sequence Data; D011728:Pyridones / Q000378:metabolism*; D012417:Rumen / Q000737:chemistry* / Q000382:microbiology
D001426:Bacterial Proteins; D007306:Insecticides; D011728:Pyridones; C012587:3,5,6-trichloro-2-pyridinol; D000581:Amidohydrolases