Production, Biochemical Characterization, and Kinetic/Thermodynamic Study of Inulinase from URM4658.

Inulinases are enzymes involved in the hydrolysis of inulin, which can be used in the food industry to produce high-fructose syrups and fructo-oligosaccharides. For this purpose, different strains and substrates were tested for inulinase production by solid-state fermentation, among which URM4658 grown on wheat bran showed the highest activity (15.08 U mL). The inulinase produced by this strain exhibited optimum activity at 60 °C and pH 4.0. A detailed kinetic/thermodynamic study was performed on the inulin hydrolysis reaction and enzyme thermal inactivation. Inulinase was shown to have a high affinity for substrate evidenced by very-low Michaelis constant values (0.78-2.02 mM), which together with a low activation energy (19.59 kJ mol), indicates good enzyme catalytic potential. Moreover, its long half-life ( = 519.86 min) and very high -value (1726.94 min) at 60 °C suggested great thermostability, which was confirmed by the thermodynamic parameters of its thermal denaturation, namely the activation energy of thermal denaturation ( = 182.18 kJ mol) and Gibbs free energy (106.18 ≤ Δ* ≤ 111.56 kJ mol). These results indicate that URM4658 inulinase is a promising and efficient biocatalyst, which could be fruitfully exploited in long-term industrial applications.