M.G. Holyavka, A.S. Belenova, E.L. Makarova, T.A. Kovaleva, V.G. Artyukhov

In the presented work there are summarized results of various researches, including own, concerning ways of regulation and activity stabilization of inulinases, lipases and glucoamylases. The special attention is paid to the characteristic of heterogeneous preparations on the basis of the immobilized enzymes and ways of their application. Having considered a large number of works and being based on the results, received in our laboratory, we were convinced, that the immobilization can lead to considerable loss of enzymes preparation activity, however, selecting the corresponding carrier and a method of formation for communication between interacting components, it is possible to reduce considerably adverse influence of a matrix on structurally functional properties of enzyme and by that to increase specific activity of the immobilized biocatalyst. It is established, that the immobilization allows to increase value of a temperature optimum (in certain cases even by 20-25 °C) for inulinases, lipases and glucoamylases functioning, stabilizes conformation of their molecules, protects spatial structure of enzymes from influence of denaturant agents. It is made the assumption, that mechanisms of stabilization for the immobilized enzyme in the conditions of extreme pH values, temperatures, and also other denaturant agents in a general way coincide and are caused, first of all, by change of mobility degree in tertiary structure of the protein, which responsible for education of enzyme-substrate complex. The scheme of processes for the adsorptive immobilization of inulinases, lipases, glucoamylases on ion-exchange resins, fibers and other agents, having on a surface positive or negative charges, for example, chitosan, is offered. By means of UR-spectroscopy it is shown, that binding of inulinases, lipases and glucoamylases with matrixes of various carriers happens generally at the expense of electrostatic interactions. It is shown, that mechanisms of interaction between these a hydrolases and matrix of cationic and anionic exchanger polymers essentially differ from each other: in the course of adsorption different sites of proteins take part, causing thereby various conformational reorganizations in an enzyme molecule. It was noted, that the universal method of hydrolases immobilization doesn't exist now, each ways has it-s advantages and disadvantages. As a whole, the choice of a method for immobilization of the biocatalyst depends on research problems and the using direction of received enzymes preparation in this or that area of science and production.