Development of an Immobilized Laccases - CDI CIMmultus® Monolithic Reactor for Ecofriendly Producing Gallic Acid-Dextran Conjugate

Enzymatic modification of polysaccharides is an efficient and environmentally friendly method to enhance their bioactivities. Immobilized enzymes represent a technology that enables the reuse of enzymes and increase their stability. In this study, laccases from Trametes versicolor were covalently immobilized on CIMmultus (R) CDI monolithic column, which was employed to graft gallic acid (GA) onto the backbone of T40 dextran. 1.43 mg laccases were successfully immobilized, yielding a mass conversion rate of 32.17 %. The kinetic parameters of free and immobilized laccases were respectively 0.0056 and 0.1770 U for Vmax and 0.0121 and 0.7510 mmol center dot l - 1 for Km. Remarkably, 91.29 % of the enzymatic activity of immobilized laccases was preserved after catalyzing 1200 ml of reactant. A central composite design was carried out to investigate the influence of T40 dextran concentration and flow rate on the phenolization. Response surface methodology was used for predicting the optimal conditions, showed that a 2.07 % mass ratio of phenolization could be achieved with 2 % dextran and a flow rate of 0.89 ml center dot min - 1 . The GA-dextran conjugate, produced under the best conditions, revealed a DPPH radical inhibition activity of 63.97 % and an IC 50 of 73.39 mu g center dot ml - 1 . This work describes a new IMER for ecofriendly producing phenolized polysaccharides.