Functional display of fungal cellulases from Trichoderma reesei on phage M13

Abstract

To test whether the phage display technology could be applied in cellulase engineering, phagemids harboring the genes encoding the mature forms of cellobiohydrolase I (CBH I) and endoglucanase I (EG I) from filamentous fungus Trichoderma reesei were constructed, respectively. CBH I and EG I fused to the phage coat protein encoded by the g3 gene were expressed and displayed on phage M13. The phage-bound cellulases retained their activities as determined by hydrolysis of the corresponding substrates, Also, their binding abilities to insoluble cellulose substrate were confirmed by an ELISA method. Overall, these results demonstrate that cellulases can be displayed on phage surface while maintaining their biological function, thus providing an alternative for directed evolution and high-throughput screening for improved cellulases.

Uses of Phage Display in Agriculture: A Review of Food-Related Protein-Protein Interactions Discovered by Biopanning over Diverse Baits

“…One avenue being explored for more efficient conversion of cellulose to glucose is through enhanced enzymatic degradation. It has been demonstrated that some cellulases and hemicellulases retain their function when fused to a viral coat protein [61, 62]. These clones can subsequently be reengineered to alter (randomize) specific regions of interest imparting novel functionalities/affinities to the displayed enzyme combinatorially.…”