Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis

In order to improve the thermostability of lipases derived from Rhizopus chinensis, we identified lipase (Lipr27RCL) mutagenesis sites that were associated with enhanced flexibility based upon B-factor analysis and multiple sequence alignment. We found that two mutated isoforms (Lipr27RCL-K64N and L...

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Autores principales: Jiang, Zhanbao, Zhang, Chengbo, Tang, Minyuan, Xu, Bo, Wang, Lili, Qian, Wen, He, Jiandong, Zhao, Zhihong, Wu, Qian, Mu, Yuelin, Ding, Junmei, Zhang, Rui, Huang, Zunxi, Han, Nanyu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Microbiology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7063977/
https://www.ncbi.nlm.nih.gov/pubmed/32194535
http://dx.doi.org/10.3389/fmicb.2020.00346
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author Jiang, Zhanbao
Zhang, Chengbo
Tang, Minyuan
Xu, Bo
Wang, Lili
Qian, Wen
He, Jiandong
Zhao, Zhihong
Wu, Qian
Mu, Yuelin
Ding, Junmei
Zhang, Rui
Huang, Zunxi
Han, Nanyu
author_facet Jiang, Zhanbao
Zhang, Chengbo
Tang, Minyuan
Xu, Bo
Wang, Lili
Qian, Wen
He, Jiandong
Zhao, Zhihong
Wu, Qian
Mu, Yuelin
Ding, Junmei
Zhang, Rui
Huang, Zunxi
Han, Nanyu
author_sort Jiang, Zhanbao
collection PubMed
description In order to improve the thermostability of lipases derived from Rhizopus chinensis, we identified lipase (Lipr27RCL) mutagenesis sites that were associated with enhanced flexibility based upon B-factor analysis and multiple sequence alignment. We found that two mutated isoforms (Lipr27RCL-K64N and Lipr27RCL-K68T) exhibited enhanced thermostability and improved residual activity, with respective thermal activity retention values of 37.88% and 48.20% following a 2 h treatment at 50°C relative to wild type Lipr27RCL. In addition, these Lipr27RCL-K64N and Lipr27RCL-K68T isoforms exhibited 2.4- and 3.0-fold increases in enzymatic half-life following a 90 min incubation at 60°C. Together these results indicate that novel mutant lipases with enhanced thermostability useful for industrial applications can be predicted based upon B-factor analysis and constructed via site-directed mutagenesis.
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spelling pubmed-70639772020-03-19 Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis Jiang, Zhanbao Zhang, Chengbo Tang, Minyuan Xu, Bo Wang, Lili Qian, Wen He, Jiandong Zhao, Zhihong Wu, Qian Mu, Yuelin Ding, Junmei Zhang, Rui Huang, Zunxi Han, Nanyu Front Microbiol Microbiology In order to improve the thermostability of lipases derived from Rhizopus chinensis, we identified lipase (Lipr27RCL) mutagenesis sites that were associated with enhanced flexibility based upon B-factor analysis and multiple sequence alignment. We found that two mutated isoforms (Lipr27RCL-K64N and Lipr27RCL-K68T) exhibited enhanced thermostability and improved residual activity, with respective thermal activity retention values of 37.88% and 48.20% following a 2 h treatment at 50°C relative to wild type Lipr27RCL. In addition, these Lipr27RCL-K64N and Lipr27RCL-K68T isoforms exhibited 2.4- and 3.0-fold increases in enzymatic half-life following a 90 min incubation at 60°C. Together these results indicate that novel mutant lipases with enhanced thermostability useful for industrial applications can be predicted based upon B-factor analysis and constructed via site-directed mutagenesis. Frontiers Media S.A. 2020-03-03 /pmc/articles/PMC7063977/ /pubmed/32194535 http://dx.doi.org/10.3389/fmicb.2020.00346 Text en Copyright © 2020 Jiang, Zhang, Tang, Xu, Wang, Qian, He, Zhao, Wu, Mu, Ding, Zhang, Huang and Han. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Jiang, Zhanbao
Zhang, Chengbo
Tang, Minyuan
Xu, Bo
Wang, Lili
Qian, Wen
He, Jiandong
Zhao, Zhihong
Wu, Qian
Mu, Yuelin
Ding, Junmei
Zhang, Rui
Huang, Zunxi
Han, Nanyu
Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title_full Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title_fullStr Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title_full_unstemmed Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title_short Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis
title_sort improving the thermostability of rhizopus chinensis lipase through site-directed mutagenesis based on b-factor analysis
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7063977/
https://www.ncbi.nlm.nih.gov/pubmed/32194535
http://dx.doi.org/10.3389/fmicb.2020.00346
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