Continuous Production of Fructose-Syrups from Inulin by Immobilized Inulinase from Recombinant Saccharomyces cerevisiae

Abstract

Recombinant exoinulinase was partially purified form the culture supernatant of S.cerevisiae by(NH4)2SO4 precipitation and PEG treatment. The purfied inulinase was immobilized onto Amino-cellulofine with glutaraldeyde as a cross-linking agent. Immobilization yield based on the enzyme activity was about 15%. Optimal pH and temperature of immobilized enzyme were found to be 5.0 and 6$0^{\circ}C$, respectively. The enzyme activity was stably maintained in the pH ranges of 4.5 to 6.0 at 6$0^{\circ}C$. 100% of enzyme activity was observed even after incubation for 24 hr at 6$0^{\circ}C$. In the operation of a packed-bed reactor containing 412U inulinase, dahalia inulin of 7.5%(w/w) concentration was completely hydrolyzed at flow rate of 2.0mL/min at 6$0^{\circ}C$, resulting in a volumetric productivity of 693 g-reducing sugars/L/h. Under the reaction conditions of 1.0mL/min flow rate with 2.5% inulin at 6$0^{\circ}C$, the reactor was successfully operated over 30 days without loss ofinulinase activity.

Keywords

• Immobilization;
• recombinant inulinase;
• fructose-syrup;
• packed-bed reactor