A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism
Background: Biodegradation is a reliable approach for efficiently eliminating persistent pollutants such as chlorpyrifos. Despite many bacteria or fungi isolated from contaminated environment and capable of degrading chlorpyrifos, limited enzymes responsible for its degradation have been identified,...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-01-01
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| Series: | Electronic Journal of Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345817300684 |
| _version_ | 1818469864902230016 |
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| author | Shuanghu Fan Kang Li Yanchun Yan Junhuan Wang Jiayi Wang Cheng Qiao Ting Yang Yang Jia Baisuo Zhao |
| author_facet | Shuanghu Fan Kang Li Yanchun Yan Junhuan Wang Jiayi Wang Cheng Qiao Ting Yang Yang Jia Baisuo Zhao |
| author_sort | Shuanghu Fan |
| collection | DOAJ |
| description | Background: Biodegradation is a reliable approach for efficiently eliminating persistent pollutants such as chlorpyrifos. Despite many bacteria or fungi isolated from contaminated environment and capable of degrading chlorpyrifos, limited enzymes responsible for its degradation have been identified, let alone the catalytic mechanism of the enzymes.
Results: In present study, the gene cpd encoding a chlorpyrifos hydrolase was cloned by analysis of genomic sequence of Paracoccus sp. TRP. Phylogenetic analysis and BLAST indicated that CPD was a novel member of organophosphate hydrolases. The purified CPD enzyme, with conserved catalytic triad (Ser155-Asp251-His281) and motif Gly-Asp-Ser-Ala-Gly, was significantly inhibited by PMSF, a serine modifier. Molecular docking between CPD and chlorpyrifos showed that Ser155 was adjacent to chlorpyrifos, which indicated that Ser155 may be the active amino acid involved in chlorpyrifos degradation. This speculation was confirmed by site-directed mutagenesis of Ser155Ala accounting for the decreased activity of CPD towards chlorpyrifos. According to the key role of Ser155 in chlorpyrifos degradation and molecular docking conformation, the nucleophilic catalytic mechanism for chlorpyrifos degradation by CPD was proposed.
Conclusion: The novel enzyme CPD was capable of hydrolyze chlorpyrifos and Ser155 played key role during degradation of chlorpyrifos. |
| first_indexed | 2024-04-13T21:30:17Z |
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| id | doaj.art-02d44531bd7348ccb291e02957f2e01e |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-04-13T21:30:17Z |
| publishDate | 2018-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-02d44531bd7348ccb291e02957f2e01e2022-12-22T02:29:11ZengElsevierElectronic Journal of Biotechnology0717-34582018-01-0131C101610.1016/j.ejbt.2017.10.009A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanismShuanghu Fan0Kang Li1Yanchun Yan2Junhuan Wang3Jiayi Wang4Cheng Qiao5Ting Yang6Yang Jia7Baisuo Zhao8Laboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaNational Institutes for Food and Drug Control, Beijing 100050, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaLaboratory of Biology, Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBackground: Biodegradation is a reliable approach for efficiently eliminating persistent pollutants such as chlorpyrifos. Despite many bacteria or fungi isolated from contaminated environment and capable of degrading chlorpyrifos, limited enzymes responsible for its degradation have been identified, let alone the catalytic mechanism of the enzymes. Results: In present study, the gene cpd encoding a chlorpyrifos hydrolase was cloned by analysis of genomic sequence of Paracoccus sp. TRP. Phylogenetic analysis and BLAST indicated that CPD was a novel member of organophosphate hydrolases. The purified CPD enzyme, with conserved catalytic triad (Ser155-Asp251-His281) and motif Gly-Asp-Ser-Ala-Gly, was significantly inhibited by PMSF, a serine modifier. Molecular docking between CPD and chlorpyrifos showed that Ser155 was adjacent to chlorpyrifos, which indicated that Ser155 may be the active amino acid involved in chlorpyrifos degradation. This speculation was confirmed by site-directed mutagenesis of Ser155Ala accounting for the decreased activity of CPD towards chlorpyrifos. According to the key role of Ser155 in chlorpyrifos degradation and molecular docking conformation, the nucleophilic catalytic mechanism for chlorpyrifos degradation by CPD was proposed. Conclusion: The novel enzyme CPD was capable of hydrolyze chlorpyrifos and Ser155 played key role during degradation of chlorpyrifos.http://www.sciencedirect.com/science/article/pii/S0717345817300684BiodegradationChlorpyrifos degradationChlorpyrifos hydrolysisEnvironmentEnzyme purificationEnzyme structureEsteraseNucleophilic catalysisOrganophosphatePersistent pollutants |
| spellingShingle | Shuanghu Fan Kang Li Yanchun Yan Junhuan Wang Jiayi Wang Cheng Qiao Ting Yang Yang Jia Baisuo Zhao A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism Electronic Journal of Biotechnology Biodegradation Chlorpyrifos degradation Chlorpyrifos hydrolysis Environment Enzyme purification Enzyme structure Esterase Nucleophilic catalysis Organophosphate Persistent pollutants |
| title | A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism |
| title_full | A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism |
| title_fullStr | A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism |
| title_full_unstemmed | A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism |
| title_short | A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism |
| title_sort | novel chlorpyrifos hydrolase cpd from paracoccus sp trp molecular cloning characterization and catalytic mechanism |
| topic | Biodegradation Chlorpyrifos degradation Chlorpyrifos hydrolysis Environment Enzyme purification Enzyme structure Esterase Nucleophilic catalysis Organophosphate Persistent pollutants |
| url | http://www.sciencedirect.com/science/article/pii/S0717345817300684 |
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