Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis
Background: To identify the critical amino acid residues that contribute to the high enzyme activity and good thermostability of Yersinia enterocolitica subsp. palearctica (Y. NSN), 15 mutants of Y. NSN were obtained by site-directed mutagenesis in this study. And their enzyme activity and thermosta...
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| Format: | Article |
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Elsevier
2016-11-01
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| Series: | Electronic Journal of Biotechnology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345816300811 |
| _version_ | 1828241897796141056 |
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| author | Yu Zhang Zhen-Hua Li Wei Zheng Zhen-Xing Tang Zhi-Liang Zhang Lu-E Shi |
| author_facet | Yu Zhang Zhen-Hua Li Wei Zheng Zhen-Xing Tang Zhi-Liang Zhang Lu-E Shi |
| author_sort | Yu Zhang |
| collection | DOAJ |
| description | Background: To identify the critical amino acid residues that contribute to the high enzyme activity and good thermostability of Yersinia enterocolitica subsp. palearctica (Y. NSN), 15 mutants of Y. NSN were obtained by site-directed mutagenesis in this study. And their enzyme activity and thermostability were assayed. Effect of several factors on the enzyme activity and thermostability of Y. NSN, was also investigated.
Results: The results showed that the I203F and D264E mutants retained approximately 75% and 70% enzyme activity, respectively, compared to the wild-type enzyme. In addition to the I203F and D264E mutants, the mutant E202A had an obvious influence on the thermostability of Y. NSN. According to the analysis of enzyme activity and thermostability of Y. NSN, we found that Glu202, Ile203 and Asp264 might be the key residues for its high enzyme activity and good thermostability.
Conclusions: Among all factors affecting enzyme activity and thermostability of Y. NSN, they failed to explain the experimental results well. One reason might be that the enzyme activity and thermostability of Y. NSN were affected not only by a single factor but also by the entire environment. |
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| format | Article |
| id | doaj.art-97cb35c78b7d4e11bd662ee37e267c1b |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-04-12T22:06:32Z |
| publishDate | 2016-11-01 |
| publisher | Elsevier |
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| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-97cb35c78b7d4e11bd662ee37e267c1b2022-12-22T03:14:54ZengElsevierElectronic Journal of Biotechnology0717-34582016-11-0124C323710.1016/j.ejbt.2016.02.010Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesisYu Zhang0Zhen-Hua Li1Wei Zheng2Zhen-Xing Tang3Zhi-Liang Zhang4Lu-E Shi5College of Life and Environmental Sciences, Hangzhou Normal University, 310016, Hangzhou, Zhejiang, ChinaCollege of Life and Environmental Sciences, Hangzhou Normal University, 310016, Hangzhou, Zhejiang, ChinaCollege of Life and Environmental Sciences, Hangzhou Normal University, 310016, Hangzhou, Zhejiang, ChinaHangzhou Tianlong Group Co. Ltd, 310021 Hangzhou, Zhejiang, ChinaCollege of Life and Environmental Sciences, Hangzhou Normal University, 310016, Hangzhou, Zhejiang, ChinaCollege of Life and Environmental Sciences, Hangzhou Normal University, 310016, Hangzhou, Zhejiang, ChinaBackground: To identify the critical amino acid residues that contribute to the high enzyme activity and good thermostability of Yersinia enterocolitica subsp. palearctica (Y. NSN), 15 mutants of Y. NSN were obtained by site-directed mutagenesis in this study. And their enzyme activity and thermostability were assayed. Effect of several factors on the enzyme activity and thermostability of Y. NSN, was also investigated. Results: The results showed that the I203F and D264E mutants retained approximately 75% and 70% enzyme activity, respectively, compared to the wild-type enzyme. In addition to the I203F and D264E mutants, the mutant E202A had an obvious influence on the thermostability of Y. NSN. According to the analysis of enzyme activity and thermostability of Y. NSN, we found that Glu202, Ile203 and Asp264 might be the key residues for its high enzyme activity and good thermostability. Conclusions: Among all factors affecting enzyme activity and thermostability of Y. NSN, they failed to explain the experimental results well. One reason might be that the enzyme activity and thermostability of Y. NSN were affected not only by a single factor but also by the entire environment.http://www.sciencedirect.com/science/article/pii/S0717345816300811Factors affecting enzyme activityNucleaseMutationMutagenesisNucleases without sequence specificity |
| spellingShingle | Yu Zhang Zhen-Hua Li Wei Zheng Zhen-Xing Tang Zhi-Liang Zhang Lu-E Shi Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis Electronic Journal of Biotechnology Factors affecting enzyme activity Nuclease Mutation Mutagenesis Nucleases without sequence specificity |
| title | Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis |
| title_full | Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis |
| title_fullStr | Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis |
| title_full_unstemmed | Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis |
| title_short | Enzyme activity and thermostability of a non-specific nuclease from Yersinia enterocolitica subsp. palearctica by site-directed mutagenesis |
| title_sort | enzyme activity and thermostability of a non specific nuclease from yersinia enterocolitica subsp palearctica by site directed mutagenesis |
| topic | Factors affecting enzyme activity Nuclease Mutation Mutagenesis Nucleases without sequence specificity |
| url | http://www.sciencedirect.com/science/article/pii/S0717345816300811 |
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