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Novel substrates for the measurement of endo-1,4-β-glucanase (endo-cellulase)

journal contribution
posted on 2014-02-19, 00:00 authored by B V McCleary, D Mangan, Robin DalyRobin Daly, S Fort, R Ivory, N McCormack
A specific and sensitive substrate for the assay of endo-1,4-β-glucanase (cellulase) has been prepared. The substrate mixture comprises benzylidene end-blocked 2-chloro-4-nitrophenyl-β-cellotrioside (BzCNPG3) in the presence of thermostable β-glucosidase. Hydrolysis by exo-acting enzymes such as β-glucosidase and exo-β-glucanase is prevented by the presence of the benzylidene group on the non-reducing end d-glucosyl residue. On hydrolysis by cellulase, the 2-chloro-4-nitrophenyl-β-glycoside is immediately hydrolysed to 2-chloro-4-nitrophenol and free d-glucose by the β-glucosidase in the substrate mixture. The reaction is terminated and colour developed by the addition of a weak alkaline solution. The assay procedure is simple to use, specific, accurate, robust and readily adapted to automation. This procedure should find widespread applications in biomass enzymology and in the specific assay of endo-1,4-β-glucanase in general.

History

Journal

Carbohydrate research

Volume

385

Pagination

9 - 17

Publisher

Elsevier

Location

Amsterdam, The Netherlands

ISSN

0008-6215

eISSN

1873-426X

Language

eng

Publication classification

C Journal article; C1.1 Refereed article in a scholarly journal

Copyright notice

2013, Elsevier