412968 A Multi-Scale Investigation of Cellulases and Their Interaction with Cellulose Allomorphs

Tuesday, November 10, 2015: 12:55 PM
258 (Salt Palace Convention Center)
Sonia K Brady1, Shishir PS Chundawat2 and Matthew J. Lang1, (1)Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, (2)Chemical Engineering and Material Science, Michigan State University, Lansing, MI

The enzymatic degradation of cellulose into monomeric sugars is a bottleneck in cellulosic biofuel production. This process is dependent not only on the individual enzymes themselves but also on the physical properties of the substrate. In this study, we use a multi-scale experimental approach to investigate the interaction of cellulolytic enzymes on both native and non-native cellulose allomorphs; specifically, reducing-end specific cellobiohydrolase (CBH I) from Trichoderma reesei and its carbohydrate-binding module (CBM). Both native cellulose I and its unnatural allomorph, cellulose III, produced using a next-generation extractive ammonia based pretreatment were used in this work. Bulk experimental measurements have revealed that CBMs have a reduced affinity for non-native cellulose allomorphs (cellulose III) compared to native cellulose (cellulose I), despite higher enzymatic hydrolysis rates. Optical tweezer based single-molecule assays are used to characterize enzyme behavior and provide a molecular-level understanding of the bulk assay results. Together, our multi-scale experiments provide insight into the cellulose degradation mechanism as a basis for engineering more efficient processes for the enzymatic degradation of cellulosic feedstocks.

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See more of this Session: Biological Conversions and Processes for Renewable Feedstocks
See more of this Group/Topical: Sustainable Engineering Forum