National Renewable Energy Laboratory, 1617 Cole Boulevard, Golden, CO 80401-3393

A -- LETTER OF INTEREST FOR CELLULASE ENZYME IMPROVEMENT PROJECT SOL DW180 POC Dr. Rafael Nieves (303) 384-6884 or David Glassner (303) 384-6820 CELLULASE ENZYME IMPROVEMENT PROJECT: The National Renewable Energy Laboratory (NREL) is seeking expressions of interest from organizations and companies interested in developing cellulase enzymes. NREL wishes to bring organizations with (1) protein engineering expertise, and (2) the ability to commercially produce improved enzymes, to this 3-year project via competitive solicitation. The anticipated funding for this project is $2 to $5 million per year, to be recompeted if necessary after the initial 3-year period. An optional information meeting will be held in Golden, CO, on September 22, 1999, to review the proposed scope of work and NREL's subcontracting procedures, and to obtain feedback from interested parties. NREL manages the Biofuels Program for the U.S. Department of Energy's Office of Fuels Development. A major project within the Biofuels Program focuses on ethanol production from cellulosic biomass. A primary objective of this project is to develop improved technologies for converting cellulosic biomass such as agricultural and forestry residues, wastepaper, municipal solid waste, and other substrates into ethanol for use as a transportation fuel. New, low-cost cellulase enzymes are needed to improve the economics and reduce the risk of investment in converting cellulosic biomass to sugars and ethanol. Cellulases with higher specific activities than current fungal cellulases are needed to meet future cost reduction objectives for biomass hydrolysis and ethanol production. The strategy is to support the development of cellulase technology that can produce the required cellulase activity for much less than the cost currently estimated. The key to this strategy is to increase the specific activity of cellulases tenfold (thus deriving more activity per gram of protein produced). This will result in a tenfold cost reduction for purchased enzyme. The specific activity of the leading commercial saccharifying celluslase system (Trichoderma reesei) is approximagely 600 FPU/g cellulase protein. A preferred characteristic of the improved cellulase enzymes is tolerance to 65 degree C. (our minimum performance target) for 4 days with less than 10% activity loss. Stability at temperatures as high as 80 degrees C. will benefit the ethanol production process. Another desired characteristic is the ability of the enzyme to completely hydrolyze the cellulose to glucose at the end product. This requires overcoming product feedback inhibition. The planned solicitation seeks to bring new technical capabilities in protein engineering to achieve the programmatic cellulase enzyme improved goals. For the purpose of the solicitation, protein engineering is defined as any combination of site-directed or random mutagenesis, gene shuffling, and enzyme/gene discovery. Parties interested in attending the meeting or receiving the request for proposal should contact Rafael Nieves at rafael_nieves@nrel.gov or (303) 384-6884 or David Glassner at (303) 384-6820 for further details and instructions. RSVP for attending the meeting is required. To RSVP, please contact Rafael Nieves at rafael_nieves@nrel.gov or (303) 384-6884. Posted 08/24/99 (W-SN371909). (0236)

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