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

Abstract 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...

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Autores principales:	Zhanping Zhou, Xiao Wang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/34a6d044f9e5469db18b04233ba482cc
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spelling	oai:doaj.org-article:34a6d044f9e5469db18b04233ba482cc2021-12-02T18:49:23ZImprove thermostability of Bacillus sp. TS chitosanase through structure-based alignment10.1038/s41598-021-95369-w2045-2322https://doaj.org/article/34a6d044f9e5469db18b04233ba482cc2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95369-whttps://doaj.org/toc/2045-2322Abstract 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.Zhanping ZhouXiao WangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zhanping Zhou Xiao Wang Improve thermostability of Bacillus sp. TS chitosanase through structure-based alignment
description	Abstract 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.
format	article
author	Zhanping Zhou Xiao Wang
author_facet	Zhanping Zhou Xiao Wang
author_sort	Zhanping Zhou
title	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_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_sort	improve thermostability of bacillus sp. ts chitosanase through structure-based alignment
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/34a6d044f9e5469db18b04233ba482cc
work_keys_str_mv	AT zhanpingzhou improvethermostabilityofbacillussptschitosanasethroughstructurebasedalignment AT xiaowang improvethermostabilityofbacillussptschitosanasethroughstructurebasedalignment
_version_	1718377553498996736

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

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