T.A. Kovaleva, M.G. Holyavka, V.G. Artyukhov, A.R. Kayumov

It is shown, that anion-exchange resin AV-17-2P doesn't essential inhibitor influence on inulinase and condition of its active center. Obviously, anion-exchanger complicates diffusion process of substrate molecules to enzymes active center and influences by that on education process of enzyme-substrate complex, reducing catalytic activity of immobilized enzyme in comparison with a soluble preparation. The assumption is put forward, that reaction of inulin hydrolysis proceeds much quicker, than substratum diffusion to functional groups of the active center of immobilized enzyme, and outside- diffusion braking leads to decrease in activity immobilized inulinase in comparison with free enzyme. It is shown, that at рН 4,0 dynamics of dependence for speed of enzymes reaction from substrate concentration practically doesn't change in comparison with optimum value рН (optimum value рН for free and immobilized inulinases is in a range 4,5-4,7); at рН 3,0 and 5,5 there is a lengthening of a pre-exponential phase of reaction. It is revealed that at inulinases immobilization on anionite AV-17-2P the рК value of catalytically important amino acids as well as for free enzyme, correspond рК carboxyl groups of asparagines and glutamines and an imidazol ring of histidine. However, linkage of enzymes molecule with a matrix of the polymeric carrier leads to shift рК the amino acids in area of great values (рК СООН groups of the free and immobilized enzymes 3,3 and 3,8 respectively, for a histidine imidazol ring - 5,5 and 5,7 respectively). On UR-spectra a heterogeneous enzyme preparation change of a form and decrease in peaks intensity 3400-3200, 3000-2100, 1255-1000 cm-1 were registered, that can testify first of all to reorganizations in protein structure, compacting of enzymes molecule and also on exiting to its surface of additional number of lateral amino groups of the lysine, arginine and histidine. Formation of more densely packed waterproof kernel in comparison with a native enzyme form is one of the reasons to decrease in catalytic activity of a preparation after an immobilization.