A simple method for determining specificity of carbohydrate-binding modules for purified and crude insoluble polysaccharide substrates

Oren Yaniv, Sadanari Jindou, Felix Frolow, Raphael Lamed, Edward A. Bayer

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

Abstract

Experimental identification of carbohydrate-binding modules (CBM) and determination of ligand specificity of each CBM are complementary and compulsory steps for their characterization. Some CBMs are very specific for their primary substrate (e.g., cellulose), whereas others are relatively promiscuous or nonspecific in their substrate preference. Here we describe a simple procedure based on in-tube adsorption of a CBM to various insoluble polysaccharides, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) for determining the distribution of the CBM between the bound and unbound fractions. This technique enables qualitative assessment of the binding strength and ligand specificity for each CBM.

Original languageEnglish
Title of host publicationBiomass Conversion
Subtitle of host publicationMethods and Protocols
EditorsMichael Himmel
Pages101-107
Number of pages7
DOIs
StatePublished - 2012

Publication series

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

Keywords

  • Binding assay
  • Carbohydrate-binding module (CBM)
  • Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE)

Fingerprint

Dive into the research topics of 'A simple method for determining specificity of carbohydrate-binding modules for purified and crude insoluble polysaccharide substrates'. Together they form a unique fingerprint.

Cite this