Tuesday, April 20, 2010
8-58

Interactions of cellulase and non-cellulase enzymes with ideal substrates and switchgrass processed by leading pretreatment technologies

Jian Shi1, Mirvat A. Ebrik1, Tim Redmond1, Bin Yang1, Charles E. Wyman1, Rebecca Garlock2, Venkatesh Balan2, Bruce E. Dale2, Venkata Ramesh Pallapolu3, Y.Y. Lee3, Youngmi Kim4, Nathan S. Mosier4, Michael R. Ladisch4, Mark T. Holtzapple5, Matthew Falls5, Rocio Sierra5, Bryon S. Donohoe6, Todd B. Vinzant7, Richard T. Elander7, Bonnie R. Hames8, Steven R. Thomas8, and Ryan E. Warner9. (1) Center for Environmental Research and Technology Department of Chemical and Environmental Engineering Bourns College of Engineer, University of California at Riverside, 1084 Columbia Avenue, Riverside, CA 92507, (2) Department of Chemical Engineering and Material Science, Michigan State University, 2527 Engineering building, East Lansing, MI 48824, (3) Chemical Engineering, Auburn University, 212 Ross Hall, Auburn, AL 36849, (4) LORRE/Ag. and Bio. Engineering, Purdue University, 500 Central Dr., West Lafayette, IN 47907, (5) Chemical Engineering, Texas A&M University, 3122 TAMU, College Station, TX 77843-3122, (6) Biosciences Center, National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, CO 80401, (7) National Bioenergy Center, National Renewable Energy Laboratory, 1617 Cole Boulevard, Golden, CO 80401, (8) Ceres, Inc., 1535 Rancho Conejo Blvd, Thousand Oaks, CA 91320, (9) Genencor, A Danisco Division, 925 Page Mill Road, Palo Alto, CA 94304

Enzymes play vital roles in hydrolyzing lignocellulosic biomass to sugars. Previous studies suggested that the accessibility and reactivity of cellulase and non-cellulase enzymes varied depending on substrates and pretreatment technologies. In this study, cellulases (Spezyme CP, beta-glucosidase, and Accelerase 1000) and non-cellulases (Multifect xylanase and pectinase, beta-xylosidase) were applied to hydrolysis of pure cellulose and xylan. Cellulases and hemicellulases resulted in distinctive sugar release patterns on Avicel, PASC and xylan. Significant amounts of xylooligomers accumulated during xylan hydrolysis with individual cellulases and hemicellulases, suggesting opportunities to optimize the enzyme cocktails. The digestibility of CAFI3 pretreated switchgrass varied with pretreatment technologies (ammonia fiber expansion, aqueous ammonia recycle, dilute sulfuric acid, sulfur dioxide impregnation, and neutral pH/liquid hot water) and were correlative to compositional changes, i.e., xylan, lignin, and acetyl removal. The adsorption capacities of cellulase and non-cellulase enzymes were also measured and correlated with initial hydrolysis rates and the physical and compositional properties of pretreated switchgrass.

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