14-27: Engineering ionic liquid tolerant cellulase enzymes

Ionic liquids (ILs) are capable of solvating biomass and separating lignin and hemicellulose from cellulose.  It is likely that (partially) dissolved cellulose could be very rapidly hydrolyzed.  Aslo,  ILs prevent contamination during the enzymatic hydrolysis step in cellulosic biofuels production.  Cellulase activity in neat IL has not been demonstrated due to the IL’s denaturing effect on the enzymes.  In this work, we searched for and found a more IL-tolerant cellulase via a mutant library generated with DNA shuffling for the purpose of hydrolysis of the cellulose in the presence of the ionic liquid (with water added).  These cellulases were also more thermo-tolerant than the wild type.   Additionally, we find that cellulases from thermophilic archaea to be more IL stable than mesophilic cellulases.