Comparative assessment of the thermostability of free and immobilised inulinase on hypercrosslinked polymers
Abstract
This study demonstrates the possibility of using heterogeneous biocatalysts in catalytic transformation reactions of inulin-containing raw materials. Binding of enzymes to carriers of different nature determines the potential for the use of immobilised enzymes as specific, regenerable, and resistant to denaturing environmental factors biocatalysts. In this study, we used styrene-divinylbenzene hypercrosslinked macroporous sorbents as carriers for the adsorption immobilisation of inulinase: cation exchangers C100H (strongly acidic) and C104 (weakly acidic) based on styrene and divinylbenzene and anion exchangers A100 (low-basic) and А500R (highly basic).
It was determined that the amount of enzyme bound to the carriers as well as the activity of immobilised inulinase depend on the concentration of hydrogen ions. It was shown that the value of the sorption parameter reaches its maximum value at pH 4.7-5.0. We observed the highest amount of sorbed protein using cation exchangers C100H (0.95 mmol/g) and C104 (0.88 mmol/g). The activity of the obtained heterogeneous biocatalysts at this level of the medium pH is 64.8-83.5% of the activity of free inulinase.
We also studied the effect of temperature (in the range of 40-70 C) on the inactivation of free and bound inulinase at the optimum pH of the medium. The kinetic parameters of the inactivation process were calculated. We determined that in the considered temperature range, the inactivation constants and activation energy of immobilised inulinase are lower compared to the native enzyme. The observed regularities suggest further study and practical use of the heterogeneous biocatalyst.
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