Xylanase covalent binding onto amidated pectin beads: Optimization, thermal, operational and storage stability studies and application.

Amidated pectin-polyethylene imine-glutaraldehyde (AP-PEI-GA) immobilizer was prepared. The ideal protocol that should be adopted during the immobilizer preparation was investigated via Box-Behnken design (BBD), and it comprised processing the AP beads with 3.4 % (w/w) PEI solution of pH 9.65 followed by 5.96 % (v/v) GA solution. The obtained AP-PEI-GA immobilizer was efficient, and it acquired 3.03 U.g-1 of immobilized xylanase (im-xylanase) activity. The computed Km and Vmax values for AP-PEI-GA im-xylanase were 16.67 mg.ml-1 and 20 g.ml-1.min-1, respectively. Through covalent coupling to AP-PEI-GA, Aspergillus niger xylanase thermodynamic properties T1/2 and D-values were increased by 2.05, 3.08, and 1.35 at 40, 50, and 60 °C, respectively. ΔHd and ΔGd for AP-PEI-GA im-xylanase at 40, 50, and 60 °C were higher than those for free form emphasizing more resistance to thermal denaturation. Im-xylanase showed 100 % activity for 20 successive cycles and hydrolyzed different agro-industrial wastes into reducing sugar and xylooligosaccharides (XOS) with more efficiency on pea peel (PP). AP-PEI-GA im-xylanase, PP weight, and hydrolysis time that should be adopted to obtain the highest reducing sugar and XOS yield were optimized through central composite design (CCD). Extracted XOS showed prebiotic and anti-oxidant activities.

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 Km and Vmax were determined to be 105.26 mg·ml-1 and 83.33 µmol·ml-1·min-1, respectively. The thermodynamics including, Ea (activation energy) and Ed (activation energy for denaturation) were determined to be 21.82 and 73.21 kJ·mol-1, Kd, T1/2, 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.