数字农科院2.0

Catalytic Efficiency Improvement in Cellobiohydrolase I by Cross-Species Domain Exchange Engineering

文献类型： 外文期刊

作者： Jing Xue;Xianzhang Jiang;Anjing Li;Jiaxin Li;Xiaoyun Su;Jianzhong Huang;Lina Qin

作者机构：

关键词： carbohydrate-binding module (CBM);Cellobiohydrolase I (CBHI);cellulose;Chaetomium thermophilum;Trichoderma reesei

期刊名称： International Journal of Molecular Sciences

ISSN： 1661-6596

年卷期： 2025 年 26 卷 9 期

页码：

收录情况： SCIE（2025版）

摘要： Understanding the molecular mechanisms of cellobiohydrolase I (CBHI), a key enzyme in cellulase complexes, is crucial for developing efficient enzymes for the degradation of lignocellulosic biomasses (LCB). Building on our previous discovery that Chaetomium thermophilum CBHI (C-CBH) exhibits significantly higher specific activity than Trichoderma reesei CBHI (T-CBH), systematic domain-swapping experiments were conducted to elucidate the structural determinants of catalytic efficiency in CBHI. Herein, the carbohydrate-binding modules (CBM) of the CBHIs from Trichoderma reesei (T-CBH) and Chaetomium thermophilum (C-CBH) were interchanged and to obtain two chimeric mutants TC-CBH and CT-CBH. These four CBHs were expressed in T. reesei, and the enzyme properties were analyzed. Comparative characterization revealed that while module exchange preserved native temperature/pH adaptability, it significantly altered substrate specificity and catalytic performance. The CT-CBH variant was identified as the most efficient biocatalyst, exhibiting four key advantages over T-CBH: (1) protein expression levels that far exceed those of T-CBH, (2) specific activity enhanced by 2.6-fold (734.5 U/μM vs. 282.5 U/μM on MU-cellobiose), (3) superior degradation capacities for filter paper (1.6-fold) and xylan, and (4) improved binding affinity for crystalline cellulose. These findings establish cross-species domain engineering as a viable strategy for creating high-performance cellulases, providing both mechanistic insights and practical solutions for lignocellulose degradation.

分类号：

  • 相关文献

[1]PlantcellwallpolysaccharidesdeconstructionbyCaldicellulosiruptorbescii. 苏小运. 2015

[2]A novel thermoacidophilic and thermostable endo-beta-1,4-glucanase from Phialophora sp G5: its thermostability influenced by a distinct beta-sheet and the carbohydrate-binding module. Zhao, Junqi,Shi, Pengjun,Huang, Huoqing,Li, Zhongyuan,Yuan, Tiezheng,Yang, Peilong,Luo, Huiying,Bai, Yingguo,Yao, Bin. 2012

[3]A novel major facilitator transporter TrSTR1 is essential for pentose utilization and involved in xylanase induction in Trichoderma reesei. Huang, Zhen-Bang,Chen, Xiu-Zhen,Qin, Li-Na,Wu, Hong-Qing,Dong, Zhi-Yang,Huang, Zhen-Bang,Su, Xiao-Yun.

[4]Overexpressing key component genes of the secretion pathway for enhanced secretion of an Aspergillus niger glucose oxidase in Trichoderma reesei. Wu, Yilan,Sun, Xianhua,Xue, Xianli,Luo, Huiying,Yao, Bin,Su, Xiaoyun,Wu, Yilan,Xie, Xiangming.

[5]Revisiting overexpression of a heterologous beta-glucosidase in Trichoderma reesei: fusion expression of the Neosartorya fischeri Bgl3A to cbh1 enhances the overall as well as individual cellulase activities. Xue, Xianli,Ma, Rui,Luo, Huiying,Su, Xiaoyun,Yao, Bin,Wu, Yilan,Qin, Xing. 2016

[6]Improving Cellulase Production In Submerged F.ermentation By The Expression O f A Vitreoscilla Hemoglobin In Trichoderma Reesei. Lin, J, Zhang, XM, Song, BR, Xue, W, Su, XY, Chen, XZ, Dong, ZY. 2017

[7]Enhanced Heterologous Expression Of Trichoderma R.eesei Cel5A/Cel6A In Pichia P astoris With Extracellular Co-Expression Of Vitreoscilla Hemoglobin. Sun, FF, Yang, HM, Bai, RH, Fang, X, Wang, F, He, J, Tu, MB. 2018

[8]Overexpressing Key Component Genes Of T.he Secretion Pathway For E nhanced Secretion Of An Aspergillus Niger Glucose Oxidase In Trichoderma Reesei. Wu, YL, Sun, XH, Xue, XL, Luo, HY, Yao, B, Xie, XM, Su, XY. 2017

[9]Exogenous microbial antagonism affects the bioaugmentation of humus formation under different inoculation using Trichoderma reesei and Phanerochaete chrysosporium. Wang L.,Sun K.,Pan S.,Wang S.,Yan Z.,Zhu L.,Yang X.. 2023

[10]Combining manipulation of integration loci and secretory pathway on expression of an Aspergillus niger glucose oxidase gene in Trichoderma reesei. Wangli Ji,Xiaolu Wang,Xiaoqing Liu,Yuan Wang,Fangui Liu,Bo Xu,Huiying Luo,Tao Tu,Wei Zhang,Xinxin Xu,Xiaoyun Su. 2023

[11]Simultaneous manipulation of multiple genes within a same regulatory stage for iterative evolution of Trichoderma reesei. Xianhua Sun,Yazhe Liang,Yuan Wang,Honglian Zhang,Tong Zhao,Bin Yao,Huiying Luo,Huoqing Huang,Xiaoyun Su. 2022

[12]Genetic engineering and raising temperature enhance recombinant protein production with the cdna1 promoter in Trichoderma reesei. Shanshan Jiang,Yue Wang,Qin Liu,Qinqin Zhao,Liwei Gao,Xin Song,Xuezhi Li,Yinbo Qu,Guodong Liu. 2022

[13]Customized optimization of lignocellulolytic enzyme cocktails for efficient conversion of pectin-rich biomass residues. Gao L.,Liu G.,Zhao Q.,Xiao Z.,Sun W.,Hao X.,Liu X.,Zhang Z.,Zhang P.. 2022

[14]Rnai-Mediated Gene Silencing Of Trcot1 Induces A Hyperbranching Phenotype In Trichoderma Reesei. Gao, F, Li, MZ, Liu, WQ, Bai, YG, Tao, T, Wang, Y, Zhang, J, Luo, HY, Yao, B, Huang, HQ, Su, XY. 2020

[15]Engineering The Cbh1 Promoter Of Trichoderma Reesei For Enhanced Protein Production By Replacing The Binding Sites Of A Transcription Repressor Ace1 To Those Of The Activators. Sun, XH, Zhang, XH, Huang, HQ, Wang, Y, Tu, T, Bai, YG, Wang, YR, Zhang, J, Luo, HY, Yao, B, Su, XY. 2020

[16]Disruption of the Trichoderma reesei gul1 gene stimulates hyphal branching and reduces broth viscosity in cellulase production. Zhao Qinqin,Liu Qin,Wang Qi,Qin Yuqi,Zhong Yaohua,Gao Liwei,Liu Guodong,Qu Yinbo. 2021

[17]A Versatile System For Fast S.creening And Isolation Of T richoderma Reesei Cellulase Hyperproducers Based On Dsred And Fluorescence-Assisted Cell Sorting. Gao, F, Hao, ZZ, Sun, XH, Qin, LN, Zhao, T, Liu, WQ, Luo, HY, Yao, B, Su, XY. 2018

[18]A closed-loop strategy for on-site production of saccharolytic enzymes for lignocellulose biorefinery using internal lignocellulosic hydrolysates. Qinqin Zhao,Zheng Zhang,Zhihao Liu,Huiqi Liang,Liwei Gao,Jian Zhao,Guodong Liu,Yinbo Qu. 2024

[19]Sub-genomic RNAi-assisted strain evolution of filamentous fungi for enhanced protein production. Sun, Xianhua,Gao, Fei,Fan, Chao,Yang, Shuyan,Zhao, Tong,Tu, Tao,Luo, Huiying,Yao, Bin,Huang, Huoqing,Su, Xiaoyun. 2024

[20]Exploiting Systematic Engineering of the Expression Cassette as a Powerful Tool to Enhance Heterologous Gene Expression in Trichoderma reesei. Ji, Wangli,Xu, Li,Sun, Xianhua,Xu, Xinxin,Zhang, Honglian,Luo, Huiying,Yao, Bin,Zhang, Wei,Su, Xiaoyun,Huang, Huoqing. 2024

作者其他论文 更多>>