Lignocellulose Recalcitrance Screening by Integrated High Throughput Hydrothermal Pretreatment and Enzymatic Saccharification

We report a novel 96-well multiplate reactor system for comparative analysis of lignocellulose recalcitrance via integrated hydrothermal pretreatment and enzymatic saccharification. The system utilizes stackable nickel/gold-plated 96-well aluminum reactor plates, a clamping device fit to a standard Parr reactor, and robotics for efficient liquids and solids handling. A capacity of 20 plates allows up to 1,920 separate hydrothermal reactions per run. Direct and rapid analysis of key end-products, glucose and xylose, is facilitated by the use of glucose oxidase/peroxidase and xylose dehydrogenase-linked assays. To demonstrate efficacy, a set of 755 poplar core samples from the US Department of Energy's BioEnergy Science Center was tested. Total sugar release ranged from 0.17 to 0.64 g/g of biomass and correlated strongly with the ratio of syringyl to guaiacyl lignins in the samples. Variance among sample replicates was sufficiently minimal to permit clear assignment of differences in recalcitrance throughout this large sample set.