PENICILLIN G ACYLASE ENZYME IMMOBILIZATION PROCESS USING A 41-4nm MCM NANOSILICA ALLOY MATRIX, Na-ALGINATE, AND CHITOSAN

Abstract

This invention relates to a process of immobilization of penicillin G acylase (PGA) enzyme using 41-4nm MCM silica nanomaterial combined with Na-alginate and chitosan, through the stages of Na-alginate 1%, 1.25%, and 1.50% (w / v) dissolved in 50 mM Tris-Cl buffer pH 7.0 at 50 °C, added 0%, 0.5%, 1.0%, and 2.0% (w / v) MCM 41-4nm nanosilica. The mixture was cooled to 22-30 °C, added 10% (v / v) PGA until well mixed. The mixture was dropped into a 2.5% (w / v) chitosan solution in 2% acetic acid and 3 M CaCl2, stirred for 2 hours at 4 °C. The formed beads were soaked in 2.5% (v / v) glutaraldehyde for 1.5 hours, then separated, washed with distilled water twice, stored in 50 mM Tris-Cl buffer pH 7.0. The use of Na-alginate resulted in a yield of immobilized PGA activity of 67.14%-87.20%. The addition of 41-4nm MCM nanosilica increased the yield of immobilized PGA activity to 87.13%-90.40%.The hydrolysis activity of immobilized PGA ranged from 2.3-7.3 U / g beads, with an optimal reaction time of 35 minutes. Soaking PGA beads in 2.5% (v / v) glutaraldehyde increased the hydrolysis activity of immobilized PGA to 9.10-14.50 U / g beads. The results of SEM analysis showed that the surface structure characteristics of immobilized PGA beads were more porous and the aggregates between the matrices were more compact.