Colorimetric Detection of Acetyl Xylan Esterase Activities

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1588)

Abstract

Colorimetric detection of reaction products is typically preferred for initial surveys of acetyl xylan esterase (AcXE) activity. This chapter will describe common colorimetric methods, and variations thereof, for measuring AcXE activities on commercial, synthesized, and natural substrates. Whereas assays using pNP-acetate, α-naphthyl acetate, and 4-methylumbelliferyl acetate (4MUA) are emphasized, common methods used to measure AcXE activity towards carbohydrate analogs (e.g., acetylated p-nitrophenyl β-d-xylopyranosides) and various acetylated xylans are also described. Strengths and limitations of the colorimetric assays are highlighted.

Key words

Acetyl xylan esterase Colorimetric assays pNP-acetate α-Naphthyl acetate 4-Methylumbelliferyl acetate Acetylated xylooligosaccharides β-Xylosidase-coupled assay 

Notes

Acknowledgments

This work was supported by a grant to G.M. from Ella and Georg Ehrnrooth foundation, Finland and an ERC Consolidator Grant to E.M. (BHIVE—648925). We thank Professor M. Tenkanen for her critical review of the manuscript.

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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Bioproducts and BiosystemsAalto UniversityEspoo, AaltoFinland
  2. 2.Department of Chemical Engineering and Applied ChemistryUniversity of TorontoTorontoCanada

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