(374aj) Microscale Platforms for High-Throughput Lignocellulosic Biomass Assays

Authors: 
Bharadwaj, R., Sandia National Laboratories, Joint BioEnergy Institute
Singh, A. K., Sandia National Laboratories
Wong, A., Sandia National Laboratories
Singh, S., Joint BioEnergy Institute
Holmes, B., The Joint BioEnergy Institute
Simmons, B. A., Joint BioEnergy Institute
Adams, P. D., The Joint BioEnergy Institute
Auer, M., The Joint BioEnergy Institute
Knierim, B., The Joint BioEnergy Institute


The high cost of lignocellulolytic enzymes is one of the main barriers towards the development of economically competitive biorefineries. Enzyme engineering can be used to significantly increase the production rate as well as specific activity of enzymes. However, the success of enzyme optimization efforts is currently limited by a lack of robust high-throughput (HTP) cellulase screening platforms for insoluble pretreated lignocellulosic substrates. We have developed a cost-effective microscale HTP enzyme-screening platform for ionic liquid (IL) pretreated lignocellulose. By performing in-situ biomass regeneration in micro-volumes, we can volumetrically meter biomass (sub-mg loading) and also precisely control the amount of residual IL for engineering novel IL-tolerant cellulases. Further, we have developed a microfluidic capillary electrophoresis device for rapid, precise, and high-throughput characterization of glycans for enzyme screening. The run-time per sample is around 60 s (> 10X faster than HPLC) and the detection sensitivity is around 1 amol. The proposed method can be used to screen individual cellulases as well as to develop novel cellulase cocktails.