Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge
Abstract Background The metagenome contains plenty of genetic resources and can be used to search for the novel gene or mutant. Results In this study, the bacterial laccase gene (cueO) with single or multiple mutations was directly cloned based on the metagenome of a chemical plant sludge. An intere...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2017-11-01
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| Series: | Bioresources and Bioprocessing |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40643-017-0178-0 |
| _version_ | 1811211130423476224 |
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| author | Qingxia Yue Yunxia Yang Jintong Zhao Lijie Zhang Li Xu Xiaoyu Chu Xiaoqing Liu Jian Tian Ningfeng Wu |
| author_facet | Qingxia Yue Yunxia Yang Jintong Zhao Lijie Zhang Li Xu Xiaoyu Chu Xiaoqing Liu Jian Tian Ningfeng Wu |
| author_sort | Qingxia Yue |
| collection | DOAJ |
| description | Abstract Background The metagenome contains plenty of genetic resources and can be used to search for the novel gene or mutant. Results In this study, the bacterial laccase gene (cueO) with single or multiple mutations was directly cloned based on the metagenome of a chemical plant sludge. An interesting mutation (G276R) was identified from those cloned mutants. The other mutants (G276N, G276Y, and G276K) with improved catalytic efficiency were identified by the saturation mutagenesis on residue G276. The optimal temperature for wild-type CueO enzyme activity was about 70 °C, compared to 60 °C, 50 °C, 50 °C, and 30 °C for the G276R, G276N, G276Y, and G276K mutant enzymes, respectively. The catalytic efficiency (k cat/K m) with 8 mmol Cu2+ of the G276R, G276N, G276Y, and G276K mutants was 1.2-, 2.7-, 1.3-, and 2.7-fold, respectively, compared to the wild-type enzyme. In addition, the mutants G276R, G276N, G276Y, and G276K oxidized the carcinogen benzo[α]pyrene more efficiently compared to the wild-type enzyme. Conclusion All of the results indicate that G276 of CueO plays an important role in enzyme activity, and the useful mutants can be identified based on the metagenome. |
| first_indexed | 2024-04-12T05:07:31Z |
| format | Article |
| id | doaj.art-d62a4c1c863f49558df86e433ff9da5c |
| institution | Directory Open Access Journal |
| issn | 2197-4365 |
| language | English |
| last_indexed | 2024-04-12T05:07:31Z |
| publishDate | 2017-11-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Bioresources and Bioprocessing |
| spelling | doaj.art-d62a4c1c863f49558df86e433ff9da5c2022-12-22T03:46:50ZengSpringerOpenBioresources and Bioprocessing2197-43652017-11-01411810.1186/s40643-017-0178-0Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludgeQingxia Yue0Yunxia Yang1Jintong Zhao2Lijie Zhang3Li Xu4Xiaoyu Chu5Xiaoqing Liu6Jian Tian7Ningfeng Wu8College of Life Science and Technology, Jinan UniversityInstitute of Quality Standards and Testing Technology for Agro-products, Chinese Academy of Agricultural SciencesBiotechnology Research Institute, Chinese Academy of Agricultural SciencesBiotechnology Research Institute, Chinese Academy of Agricultural SciencesEnvironmental Protection Research Institute of Light IndustryBiotechnology Research Institute, Chinese Academy of Agricultural SciencesBiotechnology Research Institute, Chinese Academy of Agricultural SciencesBiotechnology Research Institute, Chinese Academy of Agricultural SciencesBiotechnology Research Institute, Chinese Academy of Agricultural SciencesAbstract Background The metagenome contains plenty of genetic resources and can be used to search for the novel gene or mutant. Results In this study, the bacterial laccase gene (cueO) with single or multiple mutations was directly cloned based on the metagenome of a chemical plant sludge. An interesting mutation (G276R) was identified from those cloned mutants. The other mutants (G276N, G276Y, and G276K) with improved catalytic efficiency were identified by the saturation mutagenesis on residue G276. The optimal temperature for wild-type CueO enzyme activity was about 70 °C, compared to 60 °C, 50 °C, 50 °C, and 30 °C for the G276R, G276N, G276Y, and G276K mutant enzymes, respectively. The catalytic efficiency (k cat/K m) with 8 mmol Cu2+ of the G276R, G276N, G276Y, and G276K mutants was 1.2-, 2.7-, 1.3-, and 2.7-fold, respectively, compared to the wild-type enzyme. In addition, the mutants G276R, G276N, G276Y, and G276K oxidized the carcinogen benzo[α]pyrene more efficiently compared to the wild-type enzyme. Conclusion All of the results indicate that G276 of CueO plays an important role in enzyme activity, and the useful mutants can be identified based on the metagenome.http://link.springer.com/article/10.1186/s40643-017-0178-0Bacterial laccaseCueOG276KMutationMetagenome |
| spellingShingle | Qingxia Yue Yunxia Yang Jintong Zhao Lijie Zhang Li Xu Xiaoyu Chu Xiaoqing Liu Jian Tian Ningfeng Wu Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge Bioresources and Bioprocessing Bacterial laccase CueO G276K Mutation Metagenome |
| title | Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge |
| title_full | Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge |
| title_fullStr | Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge |
| title_full_unstemmed | Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge |
| title_short | Identification of bacterial laccase cueO mutation from the metagenome of chemical plant sludge |
| title_sort | identification of bacterial laccase cueo mutation from the metagenome of chemical plant sludge |
| topic | Bacterial laccase CueO G276K Mutation Metagenome |
| url | http://link.springer.com/article/10.1186/s40643-017-0178-0 |
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