Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR
Background: Lipases are extensively exploited in lots of industrial fields; cold-adapted lipases with alkali-resistance are especially desired in detergent industry. Penicillium cyclopium lipase I (PCL) might be suitable for applications of detergent industry due to its high catalytic efficiency at...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2019-05-01
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| Series: | Electronic Journal of Biotechnology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345819300181 |
| _version_ | 1828521383909392384 |
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| author | Lin Huang Dong Zheng Yatong Zhao Jieying Ma Yanzhen Li Zehua Xu Mengying Shan Shulin Shao Qingwen Guo Jie Zhang Fuping Lu Yihan Liu |
| author_facet | Lin Huang Dong Zheng Yatong Zhao Jieying Ma Yanzhen Li Zehua Xu Mengying Shan Shulin Shao Qingwen Guo Jie Zhang Fuping Lu Yihan Liu |
| author_sort | Lin Huang |
| collection | DOAJ |
| description | Background: Lipases are extensively exploited in lots of industrial fields; cold-adapted lipases with alkali-resistance are especially desired in detergent industry. Penicillium cyclopium lipase I (PCL) might be suitable for applications of detergent industry due to its high catalytic efficiency at low temperature and relatively good alkali stability. In this study, to better meet the requirements, the alkali stability of PCL was further improved via directed evolution with error-prone PCR. Results: The mutant PCL (N157F) with an improved alkali stability was selected based on a high-throughput activity assay. After incubating at pH 11.0 for 120 min, N157F retained 70% of its initial activity, which was 23% higher than that of wild type PCL. Combined with the three-dimensional structure analysis, N157F exhibited an improved alkali stability under the high pH condition due to the interactions of hydrophilicity and β-strand propensity. Conclusions: This work provided the theoretical foundation and preliminary data for improving alkali stability of PCL to meet the industrial requirements, which is also beneficial to improving alkali-tolerance ability of other industrial enzymes via molecular modification.How to cite: Huang L, Zheng D, Zhao Y, et al. Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR. Electron J Biotechnol 2019;39. https://doi.org/10.1016/j.ejbt.2019.04.002 Keywords: Alkali stability, Bacterial lipases, Biocatalysts, Detergent industry, Directed evolution, Enzymatic detergent, Error-prone PCR, Industrial enzymes, Penicillium cyclopium lipase |
| first_indexed | 2024-12-11T19:47:53Z |
| format | Article |
| id | doaj.art-f5ea2841266a4a499f6f40cec0dca3c4 |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-11T19:47:53Z |
| publishDate | 2019-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-f5ea2841266a4a499f6f40cec0dca3c42022-12-22T00:52:51ZengElsevierElectronic Journal of Biotechnology0717-34582019-05-01399197Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCRLin Huang0Dong Zheng1Yatong Zhao2Jieying Ma3Yanzhen Li4Zehua Xu5Mengying Shan6Shulin Shao7Qingwen Guo8Jie Zhang9Fuping Lu10Yihan Liu11State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR ChinaShandong Lonct Enzymes Co., Ltd, Shandong Province 276400, PR ChinaShandong Lonct Enzymes Co., Ltd, Shandong Province 276400, PR ChinaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China; Corresponding authors.State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China; Corresponding authors.Background: Lipases are extensively exploited in lots of industrial fields; cold-adapted lipases with alkali-resistance are especially desired in detergent industry. Penicillium cyclopium lipase I (PCL) might be suitable for applications of detergent industry due to its high catalytic efficiency at low temperature and relatively good alkali stability. In this study, to better meet the requirements, the alkali stability of PCL was further improved via directed evolution with error-prone PCR. Results: The mutant PCL (N157F) with an improved alkali stability was selected based on a high-throughput activity assay. After incubating at pH 11.0 for 120 min, N157F retained 70% of its initial activity, which was 23% higher than that of wild type PCL. Combined with the three-dimensional structure analysis, N157F exhibited an improved alkali stability under the high pH condition due to the interactions of hydrophilicity and β-strand propensity. Conclusions: This work provided the theoretical foundation and preliminary data for improving alkali stability of PCL to meet the industrial requirements, which is also beneficial to improving alkali-tolerance ability of other industrial enzymes via molecular modification.How to cite: Huang L, Zheng D, Zhao Y, et al. Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR. Electron J Biotechnol 2019;39. https://doi.org/10.1016/j.ejbt.2019.04.002 Keywords: Alkali stability, Bacterial lipases, Biocatalysts, Detergent industry, Directed evolution, Enzymatic detergent, Error-prone PCR, Industrial enzymes, Penicillium cyclopium lipasehttp://www.sciencedirect.com/science/article/pii/S0717345819300181 |
| spellingShingle | Lin Huang Dong Zheng Yatong Zhao Jieying Ma Yanzhen Li Zehua Xu Mengying Shan Shulin Shao Qingwen Guo Jie Zhang Fuping Lu Yihan Liu Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR Electronic Journal of Biotechnology |
| title | Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR |
| title_full | Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR |
| title_fullStr | Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR |
| title_full_unstemmed | Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR |
| title_short | Improvement of the alkali stability of Penicillium cyclopium lipase by error-prone PCR |
| title_sort | improvement of the alkali stability of penicillium cyclopium lipase by error prone pcr |
| url | http://www.sciencedirect.com/science/article/pii/S0717345819300181 |
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