Affinity electrophoresis as a method for determining substrate-binding specificity of carbohydrate-active enzymes for soluble polysaccharides

Sarah Moraïs, Raphael Lamed, Edward A. Bayer

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

Abstract

Affinity electrophoresis is a simple and rapid tool for the analysis of protein-binding affinities to soluble polysaccharides. This approach is particularly suitable for the characterization of the carbohydrate-active enzymes that contain a carbohydrate-binding module and for their mutants and chimeras. Knowledge of the binding characteristics of these enzymes can be the first step to elucidate the enzymatic activity of a putative enzyme; moreover in some cases, enzymes are able to bind polysaccharides targets other than their specified substrate, and this knowledge can be essential to understand the basics of the intrinsic mechanism of these enzymes in their natural environment.

Original languageEnglish
Title of host publicationBiomass Conversion
Subtitle of host publicationMethods and Protocols
EditorsMichael Himmel
Pages119-127
Number of pages9
DOIs
StatePublished - 15 Aug 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume908
ISSN (Print)1064-3745

Keywords

  • Carbohydrate-active enzymes
  • Carbohydrate-binding module (CBM)
  • Non-denaturating affinity electrophoresis
  • Substrate binding

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