BIOCATALYSIS OF CHLOROPHYLLASE IN CANOLA OIL USING ORGANIC SOLVENT SYSTEMS

Abstract

The hydrolytic activity of a partially purified chlorophyllase, obtained from an algal source, was investigated in a refined-bleached-deodorized (RBD) canola oil model system using various organic solvent media, including water/miscible-organic, biphasic organic, and micellar ternary systems. Although the use of a a ternary micellar system containing Span 85 was found to be the most appropriate medium for the hydrolysis of chlorophyll, the hydrolytic activity of chlorophyllase declined as oil content increased. The effects of various parameters, including surfactant concentration, hexane/buffer proportion, enzyme content, reaction time, incubation temperature, shaker speed, and activator concentration, on chlorophyllase activity in the micellar ternary systems containing 20% RBD oil were also investigated. In addition, phytol displayed a noncompetitive inhibition in reaction media containing polysorbate 80 and Span 85. The plot of 1/v versus oil concentrations revealed that the oil had K i values of3. 73 and 4.56% in the micellar systems containing polysorbate 80 and Span 85, respectively. Moreover, the presence of 10% oil in the micellar system containing span 85 and polysorbate 80 decreased V max values of chlorophyllase activity by 6.2 and 9.6-fold, respectively.