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Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment

Chitosanases can catalyze the release of chitooligosaccharides which have a number of medical applications. Therefore, Chitosanases are good candidates for large-scale enzymatic synthesis due to their favorable thermostability properties and high catalytic efficiency. To further improve the thermost...

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Autores principales: Zhou, Zhanping, Wang, Xiao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339078/
https://www.ncbi.nlm.nih.gov/pubmed/34349190
http://dx.doi.org/10.1038/s41598-021-95369-w
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author Zhou, Zhanping
Wang, Xiao
author_facet Zhou, Zhanping
Wang, Xiao
author_sort Zhou, Zhanping
collection PubMed
description Chitosanases can catalyze the release of chitooligosaccharides which have a number of medical applications. Therefore, Chitosanases are good candidates for large-scale enzymatic synthesis due to their favorable thermostability properties and high catalytic efficiency. To further improve the thermostability of a chitosanase from Bacillus sp. TS, which has a half-life of 5.32 min, we mutated specific serine residues that we identified as potentially relevant through structure comparison with thermophilic CelA from Clostridium thermocellum. Out of a total of 15 mutants, three, namely S265G, S276A, and S347G, show higher thermostability. Their half-lives at 60 °C were calculated as 34.57 min, 36.79 min and 7.2 min. The K(m) values of S265G, S276A and S347G mutants show substrate binding ability comparable to that of the wild-type enzyme, while the S265G mutant displays a significant decrease of enzymatic activities. Additionally, we studied the synergistic effects of combined mutations, observing that all double mutants and the triple mutant are more stable than the wild-type enzyme and single mutants. Finally, we investigated the mechanisms which might give a reasonable explanation for the improved thermostability via comparative analysis of the resulting 3D structures.
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spelling pubmed-83390782021-08-06 Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment Zhou, Zhanping Wang, Xiao Sci Rep Article Chitosanases can catalyze the release of chitooligosaccharides which have a number of medical applications. Therefore, Chitosanases are good candidates for large-scale enzymatic synthesis due to their favorable thermostability properties and high catalytic efficiency. To further improve the thermostability of a chitosanase from Bacillus sp. TS, which has a half-life of 5.32 min, we mutated specific serine residues that we identified as potentially relevant through structure comparison with thermophilic CelA from Clostridium thermocellum. Out of a total of 15 mutants, three, namely S265G, S276A, and S347G, show higher thermostability. Their half-lives at 60 °C were calculated as 34.57 min, 36.79 min and 7.2 min. The K(m) values of S265G, S276A and S347G mutants show substrate binding ability comparable to that of the wild-type enzyme, while the S265G mutant displays a significant decrease of enzymatic activities. Additionally, we studied the synergistic effects of combined mutations, observing that all double mutants and the triple mutant are more stable than the wild-type enzyme and single mutants. Finally, we investigated the mechanisms which might give a reasonable explanation for the improved thermostability via comparative analysis of the resulting 3D structures. Nature Publishing Group UK 2021-08-04 /pmc/articles/PMC8339078/ /pubmed/34349190 http://dx.doi.org/10.1038/s41598-021-95369-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Zhou, Zhanping
Wang, Xiao
Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title_full Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title_fullStr Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title_full_unstemmed Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title_short Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
title_sort improve thermostability of bacillus sp. ts chitosanase through structure-based alignment
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339078/
https://www.ncbi.nlm.nih.gov/pubmed/34349190
http://dx.doi.org/10.1038/s41598-021-95369-w
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