1B-05

 

Improved Cellulases for Biomass Conversion

 


Colin Mitchinson

 

 

Genencor International Inc.

925 Page Mill Road

Palo Alto, CA 94304

 

Phone:  (650)846 5853

Fax:  (650)845 6510

 

 

 

Genencor International has been working under a DOE subcontract for cellulase cost reduction for biomass conversion to fermentable sugars.  The goal of this three-year program was to reduce the cost of cellulase by ten-fold.  Reaching this aggressive target required improvements in both the production, and in the specific performance, of the cellulase mixture. Cellulolytic biomass conversion is performed in nature by a complex mixture of enzymes. Cellobiohydrolases play a key role and all effective cellulase mixtures contain a large excess of cellobiohydrolases over endoglucanases, suggesting that it is the exocellulase activity that is limiting.  The fundamental dependence of reaction rate on temperature predicts that large increases in performance, and decreased enzyme cost, would be achieved if the enzymatic conversion could be operated at elevated temperatures. However, T. reesei CBHI is not expected to have sufficient stability to perform at high temperatures. Therefore the thermal stability of the CBH1 enzyme from T. reesei needs to be improved. This talk discusses how we have produced CBHI enzymes with altered thermal stability and specific activity: We will present our natural diversity, site-directed mutagenesis and protein engineering / directed evolution approaches which have resulted in greatly stabilised CBHI homologs and variants.