Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei
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...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2023-12-01
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| Series: | Synthetic and Systems Biotechnology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405805X23000923 |
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| author | Qinqin Zhao Zezheng Yang Ziyang Xiao Zheng Zhang Jing Xing Huiqi Liang Liwei Gao Jian Zhao Yinbo Qu Guodong Liu |
| author_facet | Qinqin Zhao Zezheng Yang Ziyang Xiao Zheng Zhang Jing Xing Huiqi Liang Liwei Gao Jian Zhao Yinbo Qu Guodong Liu |
| author_sort | Qinqin Zhao |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-08T18:43:15Z |
| format | Article |
| id | doaj.art-c1cbc0a950d946fd949c184ce7335ae7 |
| institution | Directory Open Access Journal |
| issn | 2405-805X |
| language | English |
| last_indexed | 2024-03-08T18:43:15Z |
| publishDate | 2023-12-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Synthetic and Systems Biotechnology |
| spelling | doaj.art-c1cbc0a950d946fd949c184ce7335ae72023-12-29T04:45:40ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2023-12-0184732740Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reeseiQinqin Zhao0Zezheng Yang1Ziyang Xiao2Zheng Zhang3Jing Xing4Huiqi Liang5Liwei Gao6Jian Zhao7Yinbo Qu8Guodong Liu9State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaTaishan College, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaTobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao, 266101, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, China; Taishan College, Shandong University, Qingdao, 266237, China; Corresponding author. State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, China.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.http://www.sciencedirect.com/science/article/pii/S2405805X23000923XylanaseCellulaseTrichoderma reeseiTranscription factorGenome editing |
| spellingShingle | Qinqin Zhao Zezheng Yang Ziyang Xiao Zheng Zhang Jing Xing Huiqi Liang Liwei Gao Jian Zhao Yinbo Qu Guodong Liu Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei Synthetic and Systems Biotechnology Xylanase Cellulase Trichoderma reesei Transcription factor Genome editing |
| title | Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei |
| title_full | Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei |
| title_fullStr | Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei |
| title_full_unstemmed | Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei |
| title_short | Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei |
| title_sort | structure guided engineering of transcriptional activator xyr1 for inducer free production of lignocellulolytic enzymes in trichoderma reesei |
| topic | Xylanase Cellulase Trichoderma reesei Transcription factor Genome editing |
| url | http://www.sciencedirect.com/science/article/pii/S2405805X23000923 |
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