Sunday, May 4, 2008
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Preparation and characterization of modified cellulose substrates for cellulase digestion studies

Sunkyu Park1, Rui Katahira2, Stuart Black2, Michael E. Himmel1, and David K. Johnson1. (1) Chemical and Biosciences Center, National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, CO 80401, (2) National Bioenergy Center, National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, CO 80401

There has been an increasing interest in the conversion of lignocellulosic biomass to ethanol; however, a major barrier to commercialization of this technology is the high cost of the enzymes needed to saccharify cellulose.  A significant effort is underway in laboratories throughout the world to improve our understanding of the action of cellulolytic enzymes on the cellulose in pretreated plant cell walls.  In this study, we will demonstrate a methodology for preparing modified cellulose substrates for advanced cellulase action studies.  Cellulose substrates with different degrees of polymerization were prepared and analyzed using high performance size exclusion chromatography (HPSEC) and liquid NMR.  The method chosen for this study, carbanilation using phenyl isocyanate, minimizes sugar degradation and the formation of byproducts.  Amorphous cellulose substrates were also produced by several different methods and their properties were characterized by x-ray diffraction, NMR, differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA).  Microcrystalline celluloses and celluloses from pretreated biomass have been analyzed.  These modified cellulose substrates will be further studied with purified cellulases.

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