Enhancement of the viscometric endocellulase activity of Polyporus arcularius CMCase IIIa by cellobiose and cellooligosaccharides

Purification and viscometric characterization of three CMCases from Polyporus arcularius were carried out. The three CMCases, I, II, and IIIa, were estimated to have molecular masses of 39.1 kDa, 36.3kDa, and 24.3kDa, respectively. The addition of cellobiose and cellooligosaccharides to the reaction mixtures of CMCase I and II inhibited viscometric endocellulase activity. Following the addition of 20 mM cellobiose, CMCase I and II activities fell to about 30%-36% of their activity in the absence of cellobiose. CMCase IIIa activity, on the other hand, increased in proportion to the increase in cellobiose or cellooligosaccharide concentration. Maximal enhancement of CMCase IIIa activity was observed following the addition of cellobiose, whereas less enhancement was observed with cellooligosaccharides spanning more than two glucoside units. The addition of 20mM cellobiose resulted in an increase greater than 500% in CMCase IIIa activity. Inhibition of CMCase I and II by cellobiose and cellooligosaccharides may be the result of competition between the substrate and the reaction products. One of the reaction products of CMCase IIIa may bind to a site other than the active site of the enzyme, thus enhancing CMCase IIIa activity.