title: Cloning and expression of ophB gene encoding organophosphorus hydrolase from endophytic Pseudomonas sp. BF1-3 degrades organophosphorus pesticide chlorpyrifos.
abstract: Chlorpyrifos is an organophosphate pesticide that has adverse effect on animals and plants. We isolated endophytic bacterial strain, Pseudomonas sp. BF1-3, from balloon flower root which can hydrolyze chlorpyrifos. A gene (ophB) encoding a protein involved in chlorpyrifos degradation from this strain was cloned into Escherichia coli DH5α for confirming enzyme activity. After sequencing, total 1024bp nucleotide sequences were found in the open reading frame of ophB. The chlorpyrifos degradation patterns by E. coli DH5α (ophB) were observed. During incubation in minimal salt (M9) medium supplemented with chlorpyrifos (100mgL(-1)), the E. coli DH5α harboring ophB degraded about 97% initial chlorpyrifos (100mgL(-1)) and accumulated 86mgL(-1) 3,5,6-trichloro-2-pyridinol (TCP) within 9 days. In addition, optical density (OD) of E. coli DH5α (ophB) culture at 600nm was increased from 0.172 to 1.118 within 2 days of inoculation in the chlorpyrifos supplemented M9 medium. The estimated molecular weight of purified OphB protein was determined to be 31.4kDa by SDS-PAGE. The OphB enzyme was most active at pH 8 and an optimal temperature around 35°C. These results indicate that endophytic bacteria are supposed to be useful for biological control of environments contaminated with pesticides.
authors: Division of Applied Life Science (BK21 Plus Program), Gyeongsang National University, Chinju 660-701, Republic of Korea.
authors: Dhirendra Nath
authors: DN
authors: Barman
authors: Division of Applied Life Science (BK21 Plus Program), Gyeongsang National University, Chinju 660-701, Republic of Korea.
authors: Md Azizul
authors: MA
authors: Haque
authors: Department of Plant Pathology, Patuakhali Science and Technology University, Dumki, Patuakhali 8602, Bangladesh.
authors: Shah Md Asraful
authors: SM
authors: Islam
authors: Division of Applied Life Science (BK21 Plus Program), Gyeongsang National University, Chinju 660-701, Republic of Korea; Research Institute of Agriculture and Life Science, Gyeongsang National University, Chinju 660-701, Republic of Korea.
authors: Han Dae
authors: HD
authors: Yun
authors: Gyeongsangnam-do Agricultural Research and Extension Service, Chinju 660-360, Republic of Korea. Electronic address: goguma99@korea.kr.
authors: Min Keun
authors: MK
authors: Kim
mesh_terms: D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D001673:Biodegradation, Environmental; D004390:Chlorpyrifos / Q000378:metabolism*; D060026:Endophytes / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000254:growth & development; D007306:Insecticides / Q000378:metabolism*; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D011728:Pyridones / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D007306:Insecticides; D011728:Pyridones; D011994:Recombinant Proteins; C012587:3,5,6-trichloro-2-pyridinol; D043303:Aryldialkylphosphatase; D004390:Chlorpyrifos
keywords: Biodegradation; Chlorpyrifos; Endophytic bacteria; Organophosphorus hydrolase; Pseudomonas sp. BF1-3