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High throughput determination of glucan and xylan fractions in lignocelluloses
Authors:Michael J. Selig  Melvin P. Tucker  Cody Law  Crissa Doeppke  Michael E. Himmel  Stephen R. Decker
Affiliation:(1) Biosciences Center, National Renewable Energy Laboratory, Golden, CO 80401, USA;(2) National Bioenergy Center, National Renewable Energy Laboratory, Golden, CO 80401, USA
Abstract:The analysis of structural glucan and xylan in lignocellulose was scaled down from original two-stage sulfuric acid hydrolysis methods (Moore WE and Johnson DB 1967 Procedures for the chemical analysis of wood and wood products. U.S. Forest Products Laboratory, U.S. Department of Agriculture., Madison, WI) and integrated into a recently-developed, high throughput pretreatment and enzymatic saccharification system. Novel 96 × 1.8 ml-well Hastelloy reactor plates (128 × 86 × 51 mm) based on previously described 96-well pretreatment reactor plates were paired with custom aluminum filler plates (128 × 86 × 18 mm) for use in Symyx Powdernium solids dispensing systems. The incorporation of glucose oxidase and xylose dehydrogenase linked assays to speed post-hydrolysis sugar analysis dramatically reduced the time for analysis of large lignocellulosic sample sets. The current system permits the determination of the glucan and xylan content of 96 replicates (per reactor plate) in under 6 h and parallel plate processing increases the analysis throughput substantially.
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