Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides

Chitosanase plays an important role in enzymatic production of chitosan oligosaccharides (COSs). The present study describes the gene cloning and high-level expression of a high-efficiency chitosanase from Bacillus mojavensis SY1 (CsnBm). The gene encoding CsnBm was obtained by homologous cloning, l...

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Autores principales:	Jianrong Wang, Xiaoming Li, Hao Chen, Bilian Lin, Liangzhong Zhao
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	Bacillus mojavensis chitosanase high efficiency Pichia pastoris Chitosan oligosaccharides Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/50b30be51665414584bf1ebd54ff28ef
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spelling	oai:doaj.org-article:50b30be51665414584bf1ebd54ff28ef2021-12-01T13:53:27ZHeterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides1664-302X10.3389/fmicb.2021.781138https://doaj.org/article/50b30be51665414584bf1ebd54ff28ef2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.781138/fullhttps://doaj.org/toc/1664-302XChitosanase plays an important role in enzymatic production of chitosan oligosaccharides (COSs). The present study describes the gene cloning and high-level expression of a high-efficiency chitosanase from Bacillus mojavensis SY1 (CsnBm). The gene encoding CsnBm was obtained by homologous cloning, ligated to pPICZαA, and transformed into Pichia pastoris X33. A recombinant strain designated X33-C3 with the highest activity was isolated from 120 recombinant colonies. The maximum activity and total protein concentration of recombinant strain X33-C3 were 6,052 U/ml and 3.75 g/l, respectively, which were obtained in fed-batch cultivation in a 50-l bioreactor. The optimal temperature and pH of purified CsnBm were 55°C and 5.5, respectively. Meanwhile, CsnBm was stable from pH 4.0 to 9.0 and 40 to 55°C. The purified CsnBm exhibited high activity toward colloidal chitosan with degrees of deacetylation from 85 to 95%. Furthermore, CsnBm exhibited high efficiency to hydrolyze different concentration of colloidal chitosan to produce COSs. The result of this study not only identifies a high-efficiency chitosanase for preparation of COSs, but also casts some insight into the high-level production of chitosanase in heterologous systems.Jianrong WangJianrong WangXiaoming LiHao ChenHao ChenBilian LinBilian LinLiangzhong ZhaoLiangzhong ZhaoFrontiers Media S.A.articleBacillus mojavensischitosanasehigh efficiencyPichia pastorisChitosan oligosaccharidesMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Bacillus mojavensis chitosanase high efficiency Pichia pastoris Chitosan oligosaccharides Microbiology QR1-502
spellingShingle	Bacillus mojavensis chitosanase high efficiency Pichia pastoris Chitosan oligosaccharides Microbiology QR1-502 Jianrong Wang Jianrong Wang Xiaoming Li Hao Chen Hao Chen Bilian Lin Bilian Lin Liangzhong Zhao Liangzhong Zhao Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
description	Chitosanase plays an important role in enzymatic production of chitosan oligosaccharides (COSs). The present study describes the gene cloning and high-level expression of a high-efficiency chitosanase from Bacillus mojavensis SY1 (CsnBm). The gene encoding CsnBm was obtained by homologous cloning, ligated to pPICZαA, and transformed into Pichia pastoris X33. A recombinant strain designated X33-C3 with the highest activity was isolated from 120 recombinant colonies. The maximum activity and total protein concentration of recombinant strain X33-C3 were 6,052 U/ml and 3.75 g/l, respectively, which were obtained in fed-batch cultivation in a 50-l bioreactor. The optimal temperature and pH of purified CsnBm were 55°C and 5.5, respectively. Meanwhile, CsnBm was stable from pH 4.0 to 9.0 and 40 to 55°C. The purified CsnBm exhibited high activity toward colloidal chitosan with degrees of deacetylation from 85 to 95%. Furthermore, CsnBm exhibited high efficiency to hydrolyze different concentration of colloidal chitosan to produce COSs. The result of this study not only identifies a high-efficiency chitosanase for preparation of COSs, but also casts some insight into the high-level production of chitosanase in heterologous systems.
format	article
author	Jianrong Wang Jianrong Wang Xiaoming Li Hao Chen Hao Chen Bilian Lin Bilian Lin Liangzhong Zhao Liangzhong Zhao
author_facet	Jianrong Wang Jianrong Wang Xiaoming Li Hao Chen Hao Chen Bilian Lin Bilian Lin Liangzhong Zhao Liangzhong Zhao
author_sort	Jianrong Wang
title	Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
title_short	Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
title_full	Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
title_fullStr	Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
title_full_unstemmed	Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides
title_sort	heterologous expression and characterization of a high-efficiency chitosanase from bacillus mojavensis sy1 suitable for production of chitosan oligosaccharides
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/50b30be51665414584bf1ebd54ff28ef
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Heterologous Expression and Characterization of a High-Efficiency Chitosanase From Bacillus mojavensis SY1 Suitable for Production of Chitosan Oligosaccharides

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