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Cellobiose hydrolysis by endoglucanase (glucan glucanhydrolase) from Trichoderma reesie : Kinetics and mechanism
Author(s) -
Ladisch Michael R.,
Gong ChengShung,
Tsao George T.
Publication year - 1980
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260220602
Subject(s) - cellobiose , chemistry , trichoderma reesei , cellulase , hydrolysis , product inhibition , chromatography , kinetics , enzymatic hydrolysis , glucan , enzyme , stereochemistry , biochemistry , non competitive inhibition , physics , quantum mechanics
Glucanohydrolase from Trichoderma reesei, having a molecular weight of 52,000, was evaluated for kinetic properties with respect to cellobiose. Results from this work include: (1) initial rate studies that show that glucanohydrolase hydrolyzes cellobiose by a competitive mechanism and that the product, glucose, inhibits the enzyme; (2) low‐pressure aqueous liquid chromatography that shows that formation of a reversion product, cellobiose, is minor and occurs in detectable amounts only a very high (90m M ) cellobiose concentrations; (3) development of an equation based on the mechanism of glucanohydrolase action as determined by initial rate kinetics, which accurately predicts the time course of cellobiose hydrolysis; (4) derivation of an initial rate expression for the combined activity of cellobiase and glucanohydrolase on cellobiose. Based on data in this paper it is shown that the difference in inhibition pattern of the two enzymes could be used for determining the contamination of one enzyme by small quantities of the other.