Physiochemical, kinetic and thermodynamic studies on Aspergillus wewitschiae MN056175 inulinase with extraction of prebiotic and antioxidant Cynara scolymus leaves fructo-oligosaccharides

Utilization of agricultural wastes as cheap natural resources for production of bioactive products is currently attracting global attention. For this purpose, this study focused on isolation of Aspergillus wewitschiae MN056175 as promising producer of inulinase, then investigating physiochemical, kinetics and thermodynamics of the obtained inulinase, and its ability to extract bioactive fructo-oligosaccharides (FOS) from Cynara scolymus leaves (artichoke leaves, AL). A. wewitschiae MN056175 inulinase gave the maximum activity at temperature 60 °C and inulin concentration 1%. The kinetics including K_m and V_max were determined to be 105.26 mg·ml^-1 and 83.33 μmol·ml^-1·min^-1, respectively. The thermodynamics including, E_a (activation energy) and E_d (activation energy for denaturation) were determined to be 21.82 and 73.21 kJ·mol^-1, K_d, T_1/2, D-value, ΔH°, ΔG° and ΔS° at 40, 50 and 60 °C which indicated the stability of A. wewitschiae MN056175 inulinase. Moreover, this inulinase was capable of hydrolyzing Cynara scolymus leaves into reducing sugar and 15 FOS with different DP, total carbohydrate, and protein content under different conditions designed by central composite design (CCD). The 15 AL FOS showed different high antioxidant and prebiotic activities. Central FOS with probiotic bacteria exhibited significant antimicrobial activity against tested gram positive bacteria in a way higher than those recorded against gram negative bacteria.

Keywords: Fructo-oligosaccharides; Inulinase; Thermodynamics.