Colloid-based multiplexed method for screening plant biomass-degrading glycoside hydrolase activities in microbial communities

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The enzymatic hydrolysis of long-chain polysaccharides is a crucial step in the conversion of biomass to lignocellulosic biofuels. The identification and characterization of optimal glycoside hydrolases is dependent on enzyme activity assays, however existing methods are limited in terms of compatibility with a broad range of reaction conditions, sample complexity, and especially multiplexity. The method we present is a multiplexed approach based on Nanostructure-Initiator Mass Spectrometry (NIMS) that allowed studying several glycolytic activities in parallel under diverse assay conditions. Although the substrate analogs carried a highly hydrophobic perfluorinated tag, assays could be performed in aqueous solutions due colloid formation of ... continued below

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2884-2893

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Reindl, W.; Deng, K.; Gladden, J.M.; Cheng, G.; Wong, A.; Singer, S.W. et al. May 1, 2011.

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The enzymatic hydrolysis of long-chain polysaccharides is a crucial step in the conversion of biomass to lignocellulosic biofuels. The identification and characterization of optimal glycoside hydrolases is dependent on enzyme activity assays, however existing methods are limited in terms of compatibility with a broad range of reaction conditions, sample complexity, and especially multiplexity. The method we present is a multiplexed approach based on Nanostructure-Initiator Mass Spectrometry (NIMS) that allowed studying several glycolytic activities in parallel under diverse assay conditions. Although the substrate analogs carried a highly hydrophobic perfluorinated tag, assays could be performed in aqueous solutions due colloid formation of the substrate molecules. We first validated our method by analyzing known {beta}-glucosidase and {beta}-xylosidase activities in single and parallel assay setups, followed by the identification and characterization of yet unknown glycoside hydrolase activities in microbial communities.

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2884-2893

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  • Journal Name: Energy & Environmental Science; Journal Volume: 4; Journal Issue: 8; Related Information: Journal Publication Date: 2011

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  • Report No.: LBNL-5216E
  • Grant Number: DE-AC02-05CH11231
  • DOI: 10.1039/C1EE01112J | External Link
  • Office of Scientific & Technical Information Report Number: 1052157
  • Archival Resource Key: ark:/67531/metadc832028

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  • May 1, 2011

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  • May 19, 2016, 9:45 a.m.

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  • June 16, 2016, 1:22 p.m.

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Reindl, W.; Deng, K.; Gladden, J.M.; Cheng, G.; Wong, A.; Singer, S.W. et al. Colloid-based multiplexed method for screening plant biomass-degrading glycoside hydrolase activities in microbial communities, article, May 1, 2011; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc832028/: accessed September 20, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.