Quantification of Cellulase Activity
Quantification of active cellulases during biomass conversion processes
This technology enables the detection and quantification of active cellulases in a complex biomass mixture over time. Specific to:
- endo-b-glucanases: EG1 and EG2
- cellobiohydrolases: CBH1 and CBH2
The amount of active cellulases can be easily determined using a combination of
3 labelling agents and 3 blocking agents specific to EG1 EG2 as well as CBH1 and CBH2.
The enzymatic conversion of biopolymers to biofuels is a sophisticated way to make use of renewable energy sources. Cellulose is one of the most abundant feedstock biopolymers on earth and, unlike starch, its use is not in competition with human food sources. However, major technical challenges are associated with the conversion of cellulose to biofuels. For example, the “dying off” of cellulases during the conversion, in addition to the substrate inhomogeneity, make the control and optimization of enzyme mixture compositions during the conversion process difficult.
Researchers at The University of British Columbia have developed an activity-based protein profiling tool that allows for monitoring and quantification of the four major T. reesei model cellulases using standard separation techniques, such as SDS PAGE. The method employs synthetic fluorescence labeled probes, which irreversibly bind cellulases in their active site. Through the use of specific labeling agents and blocking agents, which inactivate specific cellulases, the activity of all four enzymes (EG1, EG2, CBH1, and CBH2) can be individually detected even in complex biomass structures and in cases where the cellulases are bound to biomass. Because this technology enables the detection and quantification of active cellulases in a complex biomass mixture over time, it is a valuable tool for control and optimization of enzyme mixture compositions, which is essential to achieve cost-effective biomass conversion.