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  • Two major facilitator superfamily sugar transporters from Trichoderma reesei and their roles in induction of cellulase biosynthesis.

Two major facilitator superfamily sugar transporters from Trichoderma reesei and their roles in induction of cellulase biosynthesis.

The Journal of biological chemistry (2013-10-03)
Weixin Zhang, Yanbo Kou, Jintao Xu, Yanli Cao, Guolei Zhao, Jing Shao, Hai Wang, Zhixing Wang, Xiaoming Bao, Guanjun Chen, Weifeng Liu
ABSTRACT

Proper perception of the extracellular insoluble cellulose is key to initiating the rapid synthesis of cellulases by cellulolytic Trichoderma reesei. Uptake of soluble oligosaccharides derived from cellulose hydrolysis represents a potential point of control in the induced cascade. In this study, we identified a major facilitator superfamily sugar transporter Stp1 capable of transporting cellobiose by reconstructing a cellobiose assimilation system in Saccharomyces cerevisiae. The absence of Stp1 in T. reesei resulted in differential cellulolytic response to Avicel versus cellobiose. Transcriptional profiling revealed a different expression profile in the Δstp1 strain from that of wild-type strain in response to Avicel and demonstrated that Stp1 somehow repressed induction of the bulk of major cellulase and hemicellulose genes. Two other putative major facilitator superfamily sugar transporters were, however, up-regulated in the profiling. Deletion of one of them identified Crt1 that was required for growth and enzymatic activity on cellulose or lactose, but was not required for growth or hemicellulase activity on xylan. The essential role of Crt1 in cellulase induction did not seem to rely on its transporting activity because the overall uptake of cellobiose or sophorose by T. reesei was not compromised in the absence of Crt1. Phylogenetic analysis revealed that orthologs of Crt1 exist in the genomes of many filamentous ascomycete fungi capable of degrading cellulose. These data thus shed new light on the mechanism by which T. reesei senses and transmits the cellulose signal and offers potential strategies for strain improvement.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cellulase from Aspergillus niger, powder, ≥0.3 units/mg solid
Sigma-Aldrich
Cellulase from Aspergillus niger, powder, off-white, ~0.8 U/mg
Sigma-Aldrich
Cellulase from Aspergillus sp., aqueous solution
Sigma-Aldrich
Cellulase from Trichoderma sp., powder, ≥5,000 units/g solid
Sigma-Aldrich
Cellulase from Trichoderma reesei, aqueous solution, ≥700 units/g
Sigma-Aldrich
Cellulase from Trichoderma sp., BioReagent, suitable for plant cell culture, 3-10 units/mg solid
Sigma-Aldrich
Cellulase from Trichoderma reesei ATCC 26921, lyophilized powder, ≥1 unit/mg solid