Enzymatic hydrolysis of Ruscus saponin in microfluidic system

In this thesis, parameters of alginate-silica hybrid monolith were optimized for microfluidic system in order to hydrolize Ruscus aculeatus extract saponin molecules and to obtain aglycones which are active compounds of some drugs. Alginate-silica hybrid beads and monoliths were obtained based on difussion gelation and internal gelation method, respectively. These hybrid beads and monoliths were characterized by SEM, EDS/mapping, BET, BJH, FTIR analyzes. Kinetic studies related with free and immobilized enzyme and their characterization studies were carried out using 4-Nitrophenyl β-D-glucopyranoside. As a result of immobilization of beta-glucosidase enzyme into alginate-silica hybrid beads a relative activity of 73% was achieved. Vmax and Km values were determined as 0.016 mmol mg-1dk-1, 4 mM and 0.012 mmol mg-1dk-1, 6 mM, respectively. Hybrid monoliths that synthesized via internal gelation technique were decided to use as a matrix. After optimize the enzyme amount (40 µg), hydrolytic conversion of pNPG were performed to understand behavior of bioreaction in microreactor. The yield in enzymatic microreactor was 20 % after first retention time, then reached the equilibrium at 24 h while the yield was 94 %. Enzymatic hydrolysis of R. aculeatus extract saponins in alginate-silica hybrid gel derived microreactor were carried out at a flow rate of 1 µl/min. Conversion yield was obtained as 78.92 % after 5 hours with 40 µg enzyme in alginate-silica derived monolithic microreactor for byproduct of R. aculeatus extract. This results showed that there is strong possibility for obtain aglycone form of saponins with further optimization of parameters in microfluidic system.