Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei  

作　　者：Qinqin Zhao Zezheng Yang Ziyang Xiao Zheng Zhang Jing Xing Huiqi Liang Liwei Gao Jian Zhao Yinbo Qu Guodong Liu 

机构地区：[1]State Key Laboratory of Microbial Technology,Shandong University,Qingdao,266237,China [2]Taishan College,Shandong University,Qingdao,266237,China [3]Tobacco Research Institute of Chinese Academy of Agricultural Sciences,Qingdao,266101,China

出　　处：《Synthetic and Systems Biotechnology》2023年第4期732-740,共9页合成和系统生物技术（英文）

基　　金：supported by the National Key R&D Program of China(2018YFA0900500);National Natural Science Foundation of China(No.32170037);the Key research program of China National Tobacco Corporation(No.110202102018)。

摘　　要：The filamentous fungus Trichoderma reesei is widely used for the production of lignocellulolytic enzymes in industry.XYR1 is the major transcriptional activator of cellulases and hemicellulases in T.reesei.However,rational engineering of XYR1 for improved lignocellulolytic enzymes production has been limited by the lack of structure information.Here,alanine 873 was identified as a new potential target for the engineering of XYR1 based on its structure predicted by AlphaFold2.The mutation of this residue to tyrosine enabled significantly enhanced production of xylanolytic enzymes in the medium with cellulose as the carbon source.Moreover,xylanase and cellulase production increased by 56.7-and 3.3-fold,respectively,when glucose was used as the sole carbon source.Under both conditions,the improvements of lignocellulolytic enzyme production were higher than those in the previously reported V821F mutant.With the enriched hemicellulases and cellulases,the crude enzymes secreted by the A873Y mutant strain produced 51%more glucose and 52%more xylose from pretreated corn stover than those of the parent strain.The results provide a novel strategy for engineering the lignocellulolytic enzyme-producing capacity of T.reesei,and would be helpful for understanding the molecular mechanisms of XYR1 regulation.

关 键 词：Xylanase Cellulase Trichoderma reesei Transcription factor Genome editing 

分 类 号：Q814[生物学—生物工程]