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Title: Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling

Abstract

Glycoside hydrolases (GHs) are enzymes that hydrolyze polysaccharides into simple sugars. To better understand the specificity of enzyme hydrolysis within the complex matrix of polysaccharides found in the plant cell wall, we studied the reactions of individual enzymes using glycome profiling, where a comprehensive collection of cell wall glycan-directed monoclonal antibodies are used to detect polysaccharide epitopes remaining in the walls after enzyme treatment and quantitative nanostructure initiator mass spectrometry (oxime-NIMS) to determine soluble sugar products of their reactions. Single, purified enzymes from the GH5_4, GH10, and GH11 families of glycoside hydrolases hydrolyzed hemicelluloses as evidenced by the loss of specific epitopes from the glycome profiles in enzyme-treated plant biomass. The glycome profiling data were further substantiated by oxime-NIMS, which identified hexose products from hydrolysis of cellulose, and pentose-only and mixed hexose-pentose products from the hydrolysis of hemicelluloses. The GH10 enzyme proved to be reactive with the broadest diversity of xylose-backbone polysaccharide epitopes, but was incapable of reacting with glucose-backbone polysaccharides. In contrast, the GH5 and GH11 enzymes studied here showed the ability to react with both glucose- and xylose-backbone polysaccharides. The identification of enzyme specificity for a wide diversity of polysaccharide structures provided by glycome profiling, and the correlatedmore » identification of soluble oligosaccharide hydrolysis products provided by oxime-NIMS, offers a unique combination to understand the hydrolytic capabilities and constraints of individual enzymes as they interact with plant biomass.« less

Authors:
 [1];  [2];  [1];  [1];  [3];  [4];  [5];  [4];  [1]
  1. Univ. of Wisconsin, Madison, WI (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  3. US Dept. of Energy Joint Bioenergy Institute, Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  4. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Georgia, Athens, GA (United States)
  5. US Dept. of Energy Joint Bioenergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1379937
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; Glycoside hydrolase; Xylanase; Xyloglucanase; Glycome profiling; Nanostructure-initiator mass spectrometry; Enzyme specificity

Citation Formats

Walker, Johnnie A., Pattathil, Sivakumar, Bergeman, Lai F., Beebe, Emily T., Deng, Kai, Mirzai, Maryam, Northen, Trent R., Hahn, Michael G., and Fox, Brian G. Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling. United States: N. p., 2017. Web. doi:10.1186/s13068-017-0703-6.
Walker, Johnnie A., Pattathil, Sivakumar, Bergeman, Lai F., Beebe, Emily T., Deng, Kai, Mirzai, Maryam, Northen, Trent R., Hahn, Michael G., & Fox, Brian G. Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling. United States. doi:10.1186/s13068-017-0703-6.
Walker, Johnnie A., Pattathil, Sivakumar, Bergeman, Lai F., Beebe, Emily T., Deng, Kai, Mirzai, Maryam, Northen, Trent R., Hahn, Michael G., and Fox, Brian G. Thu . "Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling". United States. doi:10.1186/s13068-017-0703-6. https://www.osti.gov/servlets/purl/1379937.
@article{osti_1379937,
title = {Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling},
author = {Walker, Johnnie A. and Pattathil, Sivakumar and Bergeman, Lai F. and Beebe, Emily T. and Deng, Kai and Mirzai, Maryam and Northen, Trent R. and Hahn, Michael G. and Fox, Brian G.},
abstractNote = {Glycoside hydrolases (GHs) are enzymes that hydrolyze polysaccharides into simple sugars. To better understand the specificity of enzyme hydrolysis within the complex matrix of polysaccharides found in the plant cell wall, we studied the reactions of individual enzymes using glycome profiling, where a comprehensive collection of cell wall glycan-directed monoclonal antibodies are used to detect polysaccharide epitopes remaining in the walls after enzyme treatment and quantitative nanostructure initiator mass spectrometry (oxime-NIMS) to determine soluble sugar products of their reactions. Single, purified enzymes from the GH5_4, GH10, and GH11 families of glycoside hydrolases hydrolyzed hemicelluloses as evidenced by the loss of specific epitopes from the glycome profiles in enzyme-treated plant biomass. The glycome profiling data were further substantiated by oxime-NIMS, which identified hexose products from hydrolysis of cellulose, and pentose-only and mixed hexose-pentose products from the hydrolysis of hemicelluloses. The GH10 enzyme proved to be reactive with the broadest diversity of xylose-backbone polysaccharide epitopes, but was incapable of reacting with glucose-backbone polysaccharides. In contrast, the GH5 and GH11 enzymes studied here showed the ability to react with both glucose- and xylose-backbone polysaccharides. The identification of enzyme specificity for a wide diversity of polysaccharide structures provided by glycome profiling, and the correlated identification of soluble oligosaccharide hydrolysis products provided by oxime-NIMS, offers a unique combination to understand the hydrolytic capabilities and constraints of individual enzymes as they interact with plant biomass.},
doi = {10.1186/s13068-017-0703-6},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {United States},
year = {Thu Feb 02 00:00:00 EST 2017},
month = {Thu Feb 02 00:00:00 EST 2017}
}

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Works referenced in this record:

Xylanases, xylanase families and extremophilic xylanases
journal, January 2005


Lignocellulosic Biomass Pretreatment Using AFEX
book, January 2009


Hydrolysis of lignocellulosics at low enzyme levels: Application of the AFEX process
journal, April 1996


Binding modules alter the activity of chimeric cellulases: Effects of biomass pretreatment and enzyme source
journal, July 2010

  • Kim, Tae-Wan; Chokhawala, Harshal A.; Nadler, Dana
  • Biotechnology and Bioengineering, Vol. 107, Issue 4, p. 601-611
  • DOI: 10.1002/bit.22856

A Comprehensive Toolkit of Plant Cell Wall Glycan-Directed Monoclonal Antibodies
journal, April 2010

  • Pattathil, S.; Avci, U.; Baldwin, D.
  • Plant Physiology, Vol. 153, Issue 2, p. 514-525
  • DOI: 10.1104/pp.109.151985

Deconstruction of Lignocellulosic Biomass to Fuels and Chemicals
journal, July 2011


Construction and characterization of chimeric cellulases with enhanced catalytic activity towards insoluble cellulosic substrates
journal, May 2012


Features of promising technologies for pretreatment of lignocellulosic biomass
journal, April 2005


Biosynthesis of Pectin
journal, April 2010

  • Harholt, J.; Suttangkakul, A.; Vibe Scheller, H.
  • Plant Physiology, Vol. 153, Issue 2, p. 384-395
  • DOI: 10.1104/pp.110.156588