title: Microplate-based active/inactive 1° screen for biomass degrading enzyme library purification and gene discovery.
abstract: We present here a whole-cell and permeabilized E. coli cell 1° active/inactive microplate screen for β-d-xylosidase, xylanase, endoglucanase, and ferulic acid esterase enzyme activities, which are critical for the enzymatic deconstruction of biomass for fuels and chemicals. Transformants from genomic or mutagenesis-derived libraries are screened using fluorophore-tagged substrate/enzyme activity pairs that are assayed directly in the protein expression host growth media using a minimum of specialized equipment and supplies.
authors.affiliation: USDA Agricultural Research Service, Western Regional Research Center, Albany, CA 94710, USA. kurt.wagschal@ars.usda.gov
authors.forename: Kurt
authors.initials: K
authors.lastname: Wagschal
authors.forename: Charles C
authors.initials: CC
authors.lastname: Lee
mesh_terms: D056804:Biofuels; D018533:Biomass; D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D056726:Genetic Association Studies / Q000379:methods*; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism*; D006868:Hydrolysis; D014170:Transformation, Genetic
chemical_list: D056804:Biofuels; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase; D006026:Glycoside Hydrolases