ENZYMATIC-HYDROLYSIS OF POLYSACCHARIDES IN WATER-IMMISCIBLE ORGANIC-SOLVENT, BIPHASIC SYSTEMS

Enzyme-catalyzed hydrolyzations of various polysaccharides were invest igated in two-phase, water and water-immiscible, organic solvent syste ms. Enzyme activity in water-immiscible organic solvents, such as dode cane, was strongly influenced by the amount of water present in the re action mixture. Typical bell-shaped hydrolysis curves were obtained fo r the enzymatic catalysis of agar, chitin and soluble starch, using ag arase, lysozyme and cr-amylase, respectively. Maximal hydrolysis rates for all reactions were observed at a water content of 5-10% (v/v, tot al volume). The hydrolytic conversion by alpha-amylase of various star ches to maltopentaose (G5), was greatly accelerated in the 10% (v/v, t otal volume) water content water-dodecane, biphasic system. The effect s of pH, ionic strength and water content on the time courses and rate s of the reaction in these biphasic systems were analyzed by high-perf ormance liquid chromatography (HPLC) of the products. The hydrolysis o f chitin by lysozyme was highly accelerated by a pre-treatment heating of the chitin to 90 degrees C in 10% (v/v) water/dodecane, prior to t he addition of the enzyme.