Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 68, Issue 45, Pages 12671-12682Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c04663
Keywords
transcriptional activator; Xyr1; cellulase; Bgl1; Trichoderma reesei
Funding
- National Key Research and Development Program of China [2018YFA0900500]
- National Natural Science Foundation of China [31800024, 31970029, 31770047, 31670040, 31970071]
- Major basic research projects of Natural Science Foundation of Shandong Province [ZR2019ZD19]
- Shandong Provincial Natural Science Foundation [ZR2017MC071]
- Agricultural Scientific and Technological Innovation Project of Shandong Academy of Agricultural Sciences [CXGC2018E09]
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The filamentous fungus Trichoderma reesei (teleomorph Hypocrea jecorina) is widely used as a cellulase producer in the industry. Herein, we describe the rational engineering of the publicly available T. reesei QM9414 strain to achieve a remarkable high-level production of cellulase on glucose. Overexpression of the key cellulase regulator XYR1 by the copper-repressible promoter Ptcu1 was first implemented to achieve a full cellulase production in the context of catabolite repression (CCR) while eliminating the requirement of inducing sugars for enzyme production. The T. reesei bgl1 gene was further overexpressed to compensate for its low beta-glucosidase activity on glucose. This overexpression resulted in a 102% increase in FPase activity compared with the CCR-released RUT-C30 strain cultured on Avicel. Moreover, the saccharification efficiency toward pretreated corncob residues by crude enzymes from the engineered strain on glucose increased by 85% compared with that treated by enzymes from RUT-C30 cultivated on Avicel. The engineered T. reesei strain thus shows great potential as a viable alternative to deliver commercial cellulases after further optimization for efficient saccharification of agricultural waste.
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