Cellulose Hydrolysis by Cellobiohydrolase Cel7A Shows Mixed Hyperbolic Product Inhibition

• Published in: Applied biochemistry and biotechnology 2011-09, Vol.165 (1), p.178-189
• Main Authors: Bezerra, Rui Manuel Furtado, Dias, Albino A., Fraga, Irene, Pereira, António Nazaré
• Format: Article
• Language: English
• Subjects: Biochemistry; Biological and medical sciences; Biotechnology; Cellulase; Cellulase - metabolism; Cellulose; Cellulose - metabolism; Cellulose 1,4-beta-Cellobiosidase - metabolism; Chemistry; Chemistry and Materials Science; Enzymes; Fundamental and applied biological sciences. Psychology; Fungi; Hydrolysis; Kinetics; Substrate Specificity
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-011-9242-y

In order to establish which are the contribution of linear (total), hyperbolic (partial) or parabolic inhibitions by cellobiose, and also a special case of substrate inhibition, the kinetics of cellobiohydrolase Cel7A obtained from Trichoderma reesei was investigated. Values of kinetic parameters were estimated employing integrated forms of Michaelis–Menten equations through the use of non-linear regression, and criteria for selecting inhibition models are discussed. With cellobiose added at the beginning of the reaction, it was found that cellulose hydrolysis follows a kinetic model, which takes into account a mixed hyperbolic inhibition, by cellobiose with the following parameter values: K m 5.0 mM, K ic 0.029 mM, K iu 1.1 mM, k cat 3.6 h −1 and k cat′ 0.2 h −1 . Cellulose hydrolysis without initial cellobiose added also follows the same inhibition model with similar values (4.7, 0.029 and 1.5 mM and 3.2 and 0.2 h −1 , respectively). According to Akaike information criterion, more complex models that take into account substrate and parabolic inhibitions do not increase the modulation performance of cellulose hydrolysis.