1B-04

High Throughput Discovery of Novel Cellobiohydrolases Using GigaMatrix^TM

David L. Blum*, Joslin Gemsch, Mark Dycaico, Di Wu, Irene Andruszkiewicz,

Mike Lafferty and Geoff Hazlewood

Diversa Corporation, San Diego, CA 92121

4955 Directors Place

San Diego, CA 92121

Phone: (858)526-5237

Fax: (858)526-5737

E-mail: dblum@diversa.com

Efficient utilization of lignocellulosic materials is pivotal to cost-effective generation of fuels and chemicals from biomass. In addition to endoglucanases and beta-glucosidases, a key enzyme in the hydrolysis of cellulose is cellobiohydrolase (CBH). This enzyme catalyzes the release of terminal cellobiose units from cellulose. CBHs are important because they are capable of disrupting the hydrogen-bonding between adjacent cellulose chains, thus making the substrate more available for degradation. Although there are numerous examples of this enzyme in the primary literature, the CBHs reported to date are handicapped by the fact that they have low turnover rates. The objective of this study is to discover and evolve a CBH with at least ten-fold greater specific activity against crystalline cellulose than current enzymes. GigaMatrix^TM is a proprietary high throughput screening technology that utilizes a 100,000 well format that fits into a 96 well footprint. Using this technology, new enzymes have been discovered from Diversa’s proprietary library collection with the potential for use in the conversion of lignocellulosic material to fuels and chemicals. The capacity of the enzymes discovered in this screen to efficiently degrade crystalline cellulose will be discussed as well as the potential to optimize their performance using Diversa’s evolution technologies.