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Cellobiase from Trichoderma viride : Purification, properties, kinetics, and mechanism
Author(s) -
Gong ChengShung,
Ladisch Michael R.,
Tsao George T.
Publication year - 1977
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.260190703
Subject(s) - cellobiose , trichoderma viride , cellulase , chemistry , product inhibition , hydrolysis , chromatography , kinetics , cellulose , substrate (aquarium) , enzyme , biochemistry , non competitive inhibition , biology , food science , ecology , physics , quantum mechanics
Three distinct cellobiase components were isolated from a commercial Trichoderma viride cellulase preparation by repeated chromatography on DEAE cellulose eluting by a salt gradient. The purified cellobiase preparations were evaluated for physical properties, kinetics, and mechanism. Results from this work include: (1) development of a one step enzyme purification procedure using DEAE‐cellulose; (2) isolation of three chromatographically distinct, yet kinetically similar, cellobiase fractions of molecular weight of ∼76,000; (3) determination of kinetics which shows that cellobiase hydrolyzes cellobiose by a noncompetitive mechanism and that the product, glucose, inhibits the enzyme, and (4) development of an equation, based on the mechanism of cellobiase action, which accurately predicts the time course of cellobiose hydrolysis over an eightfold range of substrate concentration and conversions of up to 90%. Based on the data presented in the paper, it is shown that product inhibition of cellobiase significantly retards the rate of cellobiose hydrolysis.