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United States Department of Agriculture

Agricultural Research Service

Title: A microplate assay for quantitative evaluation of plant cell wall degrading enzymes

Authors
item King, B.C. - CORNELL UNIVERSITY
item Donnelly, M.K. - CORNELL UNIVERSITY
item Bergstrom, G.C. - CORNELL UNIVERSITY
item Walker, L.P. - CORNELL UNIVERSITY
item Gibson, Donna

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: August 26, 2008
Publication Date: August 28, 2008
Citation: King, B., Donnelly, M., Bergstrom, G., Walker, L., Gibson, D.M. 2008. A microplate assay for quantitative evaluation of plant cell wall degrading enzymes. Meeting Abstract. p. 20.

Technical Abstract: Developing enzyme cocktails for cellulosic biomass hydrolysis complementary to current cellulase systems is a critical step needed for economically viable biofuels production. Plant pathogenic fungi are a largely untapped resource in which to prospect for novel hydrolytic enzymes for biomass conversion. A standardized microplate assay for rapid analysis of polysaccharide hydrolysis by fungal extracts was developed. Fungi were grown for 10 days on cellulose- or biomass-containing media to produce enzyme extracts for analysis. Reducing sugar released from filter paper, Avicel, corn stalk, switchgrass, carboxymethylcellulose, and arabinoxylan was quantified using a miniaturized colorimetric assay based on 3,5-dinitrosalicylic acid. Significant interactions were identified among fungal species, growth media composition, assay substrate, and temperature. Within a small sampling of plant pathogenic fungi, some extracts had crude activities comparable to or greater than T. reesei, particularly when assayed at lower temperatures and on biomass substrates. This microplate assay system should prove useful for high-throughput bioprospecting for new sources of novel enzymes for biofuel production.

Last Modified: 10/20/2014
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