An alternative method for the determination of polysaccharide cleavage enzymes activities

Abstract : Most of usual methods used to detect and quantify polysaccharide cleavage enzymes (PCE) are not adapted to high throughput. In this work, enzymatic activities of three enzymes, namely an endocellulase (EC 3.2.1.4), an exocellulase (EC 3.2.1.91) and an alginate lyase (EC 4.2.2.3) were characterized by classical Michaelian model (Km, Vmax). Enzymatic activities were then determined by the use of the Enzyme Ring Test® (ERT®) method which allowed following the depolymerization of polysaccharides by PCE. Kinetic parameters were determined and compared to the ones obtained by classical methods. Besides, the ERT® technology clearly showed the possibility to discriminate endolytic and exolytic enzymatic mechanisms.