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The use of 4‐methylumbelliferyl and other chromophoric glycosides in the study of cellulolytic enzymes
Author(s) -
van Tilbeurgh Herman,
Claeyssens Marc,
de Bruyne Clement K.
Publication year - 1982
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(82)81092-2
Subject(s) - trichoderma reesei , glycoside hydrolase , cellulase , chemistry , glycoside , substrate (aquarium) , lactose , enzyme , stereochemistry , biochemistry , chromatography , biology , ecology
HPLC‐analysis of the reaction products of a series of 4‐methylumbelliferyl glycosides from cello‐oligosaccharides, used as substrates of a cellobiohydrolase from Trichoderma reesei , proves the lack of specificity for terminal cellobiosyl groups. Also, different reaction patterns are observed for this CBHI and for an endocellulase, when acting on these same substrates. 4‐Methylumbelliferyl β‐D‐lactoside is an unexpected substrate for CBHI, yielding only lactose and phenol as reaction products. The binding characteristics of p ‐nitrobenzyl 1‐thio‐β‐D‐lactoside for this enzyme are determined by a dia‐filtration technique, yielding 1 binding site and an association constant of 4.0 × 10 4 M −1 .