Conversion of biomass to precursor sugars and alcohols is complicated by the composite structure of lignocellulose which renders targeted cellulose inaccessible behind a matrix of hemicellulose and a coating of lignin. This structure necessitates pretreatment of biomass with chemical, physical, or biological means. Biological pretreatment with enzyme formulations or “cocktails” that degrade each component holds great promise as a sustainable method; however, current efficiencies of these formulations limit the cost-competitiveness of this strategy. While the need to improve enzyme efficiency has been recognized, methods of quantitatively measuring activity are limited by time-consuming or insensitive colorimetric and viscometric assays which require specialized equipment and often only report activity indirectly. Here, we demonstrate a library of substrate-functionalized single-walled carbon nanotubes (SWNT) as modular probes for characterization of cellulase, xylanase, and lignin-modifying enzymes. We also explore the effects of substrate substitution ratios and polymer lengths on consistency and selectivity (exo- vs. endo- preference) of cellulases using carboxymethylcellulose-functionalized SWNT. Lastly, we demonstrate the utility of such probes by screening an array of enzyme formulations with respect to cellulytic, hemicellulytic, and lignin-modifying activities with a single-wavelength fluorescence detector. We also show progress towards such screening in micro-droplet reactors. These sensors will find utility as real-time activity probes for improvement of enzyme operating and formulation conditions as well as rapid-screening tools for directed enzyme evolution.

Substrate-functionalized single-walled carbon nanotubes as modular activity probes for optimization of lignocellulosic biomass pretreatment

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.