Adams, Paul D.

Bharadwaj, Rajiv

Chandrasekaran, Aarthi

Hadi, Masood Z.

Park, Joshua I.

Sapra, Rajat

Simmons, Blake A.

Singh, Anup K.

eScholarship - University of California

Lawrence Berkeley National LaboratoryLBL PublicationsTitleA Microscale Platform for Integrated Cell-Free Expression and Screening of Cellulase ActivityPermalinkhttps://escholarship.org/uc/item/5xp991cgAuthorsChandrasekaran, AarthiBharadwaj, RajivPark, Joshua I.et al.Publication Date2009-12-10eScholarship.org Powered by the California Digital LibraryUniversity of California A Microscale Platform for Integrated Cell-Free Expression and Screening of Cellulase Activity  Aarthi Chandrasekaran1, Rajiv Bharadwaj1, Joshua I. Park2, Masood Z. Hadi1, Rajat Sapra2, Blake A. Simmons2 , Paul D. Adams1, Anup K. Singh1 Presenting author: Aarthi Chandrasekaran 1Technology Division, 2Deconstruction Division, Joint BioEnergy Institute, Emeryville, CA  A critical step in the efficient conversion of lignocellulosic biomass to fuel is the deconstruction of the biomass to fermentable sugars. Several efforts are therefore focused on identification, expression and characterization of novel enzymes that hydrolyze lignocellulosic biomass. High-throughput enzyme assays that enable rapid screening of these enzymes can greatly accelerate the current enzyme engineering efforts for biofuels development. In this study, we have developed a miniaturized high-throughput, fluorescence-based screening platform for rapid activity profiling of thermophilic cellulases at elevated temperatures (>80°C). This platform integrates cell-free expression and functional characterization of the cellulases in microwell arrays with volumes as low as two microliters. Herein, we demonstrate the use of this approach to express and screen a panel of thermophilic β-glucosidases and cellobiohydrolases.  This work was part of the DOE Joint BioEnergy Institute (http://www.jbei.org) supported by the U.S. Department of Energy, Office of Science, Office of Biological and E vironmental Research, through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the U.S. Departm nt of Energy.    

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A Microscale Platform for Integrated Cell-Free Expression and Screening of Cellulase Activity

https://escholarship.org/uc/item/5xp991cg

A Microscale Platform for Integrated Cell-Free Expression and Screening of Cellulase Activity

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