The Slowdown of the Endoglucanase Trichoderma reesei Cel5A-Catalyzed Cellulose Hydrolysis Is Related to Its Initial Activity

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	The Slowdown of the Endoglucanase Trichoderma reesei Cel5A-Catalyzed Cellulose Hydrolysis Is Related to Its Initial Activity
	Shu, Zhiyu; Wang, Yefei; An, Liaoyuan; Yao, Lishan
刊名	BIOCHEMISTRY
	2014-12-09
卷号	53 期号:48 页码:7650-7658
英文摘要	One important feature of hydrolysis of cellulose by cellulases is that the reaction slows down quickly after it starts. In this work, we investigate the slowdown mechanism at the early stage of the reaction using endoglucanase Tr. Cel5A-catalyzed phosphate acid-swollen cellulose (PASC) hydrolysis as a model system. Specifically, we focus on the effect of enzyme adsorption on the reaction slowdown. Nineteen single mutations are introduced (with the assistance of molecular dynamics simulations) to perturb the enzyme PASC interaction, yielding the adsorption partitioning coefficient Kr that ranged from 0.12 to 0.39 L/g, compared to that of the wild type (0.26 L/g). Several residues, including T18, K26, Y26, H229, and T300, are demonstrated to be important for adsorption of the enzyme to PASC. The kinetic measurements show that the slowdown of the hydrolysis is not correlated with the adsorption quantified by the partitioning coefficient Kr but is anticorrelated with the initial activity. This result suggests that the mutants with higher activity are more prone to being trapped or deplete the most reactive substrate faster and the adsorption plays no apparent role in the reaction slowdown. The initial activity of Cel5A against PASC is correlated with the enzyme specific activity against a soluble substrate p-nitrophenyl cellobioside.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemistry & Molecular Biology
研究领域[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	CARBOHYDRATE-BINDING MODULES ; GROMOS FORCE-FIELD ; ENZYMATIC-HYDROLYSIS ; CRYSTAL-STRUCTURE ; INSOLUBLE CELLULOSE ; CELLOBIOSE DEHYDROGENASE ; CELLOBIOHYDROLASE CEL7A ; AROMATIC RESIDUES ; CATALYTIC CORE ; X-RAY
收录类别	SCI
语种	英语
WOS记录号	WOS:000346323600020
公开日期	2015-12-24
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/6230]
专题	青岛生物能源与过程研究所_仿真模拟团队
作者单位	Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biofuels, Qingdao 266061, Peoples R China
推荐引用方式 GB/T 7714	Shu, Zhiyu,Wang, Yefei,An, Liaoyuan,et al. The Slowdown of the Endoglucanase Trichoderma reesei Cel5A-Catalyzed Cellulose Hydrolysis Is Related to Its Initial Activity[J]. BIOCHEMISTRY,2014,53(48):7650-7658.
APA	Shu, Zhiyu,Wang, Yefei,An, Liaoyuan,&Yao, Lishan.(2014).The Slowdown of the Endoglucanase Trichoderma reesei Cel5A-Catalyzed Cellulose Hydrolysis Is Related to Its Initial Activity.BIOCHEMISTRY,53(48),7650-7658.
MLA	Shu, Zhiyu,et al."The Slowdown of the Endoglucanase Trichoderma reesei Cel5A-Catalyzed Cellulose Hydrolysis Is Related to Its Initial Activity".BIOCHEMISTRY 53.48(2014):7650-7658.