Jump to main content
Jump to site search

Issue 8, 2011
Previous Article Next Article

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

Author affiliations

Abstract

The enzymatic hydrolysis of polysaccharides into fermentable sugars is a crucial step in the conversion of biomass to lignocellulosic biofuels. An efficient hydrolysis is highly dependent on the identification and characterization of optimal glycoside hydrolases. However, existing techniques for characterizing activity are limited by the range of reaction conditions that can be used, sample complexity, and throughput. The method we present is a multiplexed approach based on nanostructure-initiator mass spectrometry (NIMS) that allows for the rapid analysis of several glycolytic activities in parallel under diverse assay conditions. By forming colloids, it was possible to perform aqueous reactions in tubes and microwell plates despite the substrate analogs' hydrophobic perfluorinated tags. This method was validated by analyzing standard enzymatic parameters (temperature, pH, and kinetics) of β-glucosidase and β-xylosidase in separate setups. The multiplexity of this assay system was demonstrated by the simultaneous analysis of β-glucosidase and β-xylosidase activities, which was then used to profile environmental samples. Enzymes secreted by microbial communities within these samples were extracted by washing the sample with buffer. Enzymatic activities were directly detected within this crude extract without any further sample pretreatment steps. The multiplexed analysis of β-glucosidase, exo-/endoglucanase, and xylanase activities was applied for a detailed characterization of previously unknown glycoside hydrolase activities. The results show the suitability of the described method for the rapid screening of crude environmental samples under a wide range of conditions to determine enzyme activities from the microbial communities present within these samples.

Graphical abstract: Colloid-based multiplexed screening for plant biomass-degrading glycoside hydrolase activities in microbial communities

Back to tab navigation

Supplementary files

Publication details

The article was received on 31 Jan 2011, accepted on 08 Jun 2011 and first published on 30 Jun 2011


Article type: Paper
DOI: 10.1039/C1EE01112J
Citation: Energy Environ. Sci., 2011,4, 2884-2893
  •   Request permissions

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

    W. Reindl, K. Deng, J. M. Gladden, G. Cheng, A. Wong, S. W. Singer, S. Singh, J.-C. Lee, C.-H. Yao, T. C. Hazen, A. K. Singh, B. A. Simmons, P. D. Adams and T. R. Northen, Energy Environ. Sci., 2011, 4, 2884
    DOI: 10.1039/C1EE01112J

Search articles by author

Spotlight

Advertisements