Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state

doi:10.1002/bit.25887

QIBEBT-IR > 蛋白质设计研究组

	Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state
	Wang, Yefei 1,2; Song, Xiangfei 1,2; Zhang, Shujun 1,2; Li, Jingwen 1,2; Shu, Zhiyu 1,2; He, Chunyan 1,2; Huang, Qingshan 1,2; Yao, Lishan 1,2
	2016-06-01
发表期刊	BIOTECHNOLOGY AND BIOENGINEERING
卷号	113 期号:6 页码:1171-1177
摘要	Trichoderma reesei (Tr.) cellulases, which convert cellulose to reducing sugars, are a promising catalyst used in the lignocellulosic biofuel production. Improving Tr. cellulases activity, though very difficult, is highly desired due to the recalcitrance of lignocellulose. Meanwhile, it is preferable to enhance the cellulase's promiscuity so that substrates other than cellulose can also be hydrolyzed. In this work, an attempt is made to improve the catalytic activity of a major endogluanase Tr. Cel7B against xylan which crosslinks with cellulose in lignocellulose. By using quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations, the transition state of the xylo-oligosaccharide hydrolysis is identified. Then, mutations are introduced and their effect on the transition state stabilization is ranked based on the free energy calculations. Seven top ranked mutants are evaluated experimentally. Three mutants A208Q, A222D, and G230R show a higher activity than the wild-type Tr. Cel7B in the hydrolysis of xylan (by up to 47%) as well as filter paper (by up to 50%). The combination of the single mutants can further improve the enzyme activity. Our work demonstrates that the free energy method is effective in engineering the Tr. Cel7B activity against xylan and cellulose, and thus may also be useful for improving the activity of other Tr. cellulases. (C) 2015 Wiley Periodicals, Inc.
文章类型	Article
关键词	Cel7b Transition State Cellulase Molecular Dynamics Simulation Protein Engineering
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1002/bit.25887
关键词[WOS]	CELLULOSE HYDROLYSIS ; CELLOBIOHYDROLASE I ; QM/MM METADYNAMICS ; CATALYTIC MECHANISM ; CRYSTAL-STRUCTURE ; ENDOGLUCANASE-I ; FREE-ENERGIES ; ENZYMES ; RESOLUTION ; COMPLEX
收录类别	SCI
语种	英语
WOS研究方向	Biotechnology & Applied Microbiology
项目资助者	National Natural Science Foundation of China(21173247 ; "135'' Projects Fund of CAS-QIBEBT Director Innovation Foundation ; 21106169 ; 31400061)
WOS类目	Biotechnology & Applied Microbiology
WOS记录号	WOS:000375119900003
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/8266
专题	蛋白质设计研究组
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Synthet Biol, Qingdao 266061, Peoples R China 2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biofuels, Qingdao 266061, Peoples R China
推荐引用方式 GB/T 7714	Wang, Yefei,Song, Xiangfei,Zhang, Shujun,et al. Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state[J]. BIOTECHNOLOGY AND BIOENGINEERING,2016,113(6):1171-1177.
APA	Wang, Yefei.,Song, Xiangfei.,Zhang, Shujun.,Li, Jingwen.,Shu, Zhiyu.,...&Yao, Lishan.(2016).Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state.BIOTECHNOLOGY AND BIOENGINEERING,113(6),1171-1177.
MLA	Wang, Yefei,et al."Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state".BIOTECHNOLOGY AND BIOENGINEERING 113.6(2016):1171-1177.