Engineered Hydrolases for Biofuel Production from Cellulosic Biomass

Cellulases with mutations to supercharge the CBM (D3, D4) degrade 2.5-fold more cellulose after 48 hours at elevated temperatures​

Invention Summary:

Cellulosic biomass is a substantial fuel source with ~200 billion tons of plant biomass produced yearly. However, releasing sugars from biomass by enzymatic degradation using Carbohydrate Active enZymes (CAZymes) is challenging due to the limited accessibility to polysaccharides within the biomass structure, non-productive binding of enzymes, and low enzyme activity on highly crystalline cellulose. Biomass recalcitrance increases the industrial processing cost, impacting the viability of biomass utilization as a sustainable fuel source.

To address this issue Rutgers researchers have used enzyme supercharging to modify cellulase catalytic domain and carbohydrate binding module (CBM) surfaces by selectively introducing positively or negatively charged amino acids in solvent-exposed areas to create highly -/+ net charges. The invention consists of a library of supercharged enzymes comprising of family 5 glycoside hydrolase (GH5) Cel5A and family 2a carbohydrate binding module (CBM) from an industrially relevant thermophilic microbe Thermobifida fusca.

Market Applications:

  • Biofuel production from lignocellulosic biomass.
  • Integrate to the current cellulase blends used in the industry to boost cocktail activity even further.


  • 2-3-fold increase in enzyme activity on crystalline cellulose and lignocellulosic biomass.
  • Improved activity at elevated temperatures for several mutants.
  • Differentially charged mutants have also shown improved activity on different substrates correlating to the enzyme net charge, allowing fine-tuning enzymes for distinct substrates.
  • Shifted pH optima optimization is readily possible for different fermentation conditions and biofuel production applications.

Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration. For any business development and other collaborative partnerships contact:

Patent Information:
Lisa Lyu
Assistant Director
Rutgers, The State University of New Jersey