Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state
文献类型：期刊论文
| 作者 | Wang, Yefei1,2; Song, Xiangfei1,2; Zhang, Shujun1,2; Li, Jingwen1,2; Shu, Zhiyu1,2; He, Chunyan1,2; Huang, Qingshan1,2; Yao, Lishan1,2 |
| 刊名 | BIOTECHNOLOGY AND BIOENGINEERING
 |
| 出版日期 | 2016-06-01 |
| 卷号 | 113期号:6页码:1171-1177 |
| 关键词 | cel7B transition state cellulase molecular dynamics simulation protein engineering |
| 英文摘要 | 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 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记录号 | WOS:000375119900003 |
| 源URL | [http://ir.qibebt.ac.cn/handle/337004/8266]  |
| 专题 | 青岛生物能源与过程研究所_仿真模拟团队 |
| 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. |
入库方式： OAI收割
来源：青岛生物能源与过程研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。