ISSN 2308-4057 (Печать),
ISSN 2310-9599 (Онлайн)


Enzyme inhibitors are widely used in experimental studies in various spheres to evaluate the mechanism of catalytic effect of enzymes, determine the nature of protein reactive groups, to identify the role of various enzymatic processes in metabolism. But inhibitors are not stable and thus, they need to be stabilized through immobilization on matrixes. The study of immobilization using the infrared spectroscopy ensures to prove the interaction between the inhibitor and polysaccharidic matrix. The results of infrared spectroscopy showed that the linking between the matrix and the inhibitor occurs by formation of intramolecular covalent linkings, electrostatic correlation between the charged groups of agar and inhibitor. The derived comparison curve VVOP shows the reduction in the intensity of the immobilized inhibitor (dietar y supplement) in the area of 3400 cm-1, that is consistent with the valent variations of the free group -ОН that indicates on strengthening of the immobilized specimen hydrogen binding. Comparative study of рН-optimum of pancreatic α-amylase, native and immobilized inhibitor made it possible to conclude that рН-optimum of pancreatic α-amylase isрН 6.0, that of the native inhibitor α-amylase equals to 5.5, and the рН-optimum in the immobilized inhibitor considerably varies from 5.0 to 6.8 at the physiologic temperature (37 ± 1)ºС. Linearization methods of Michaelis-Menten equation by Lineweaver-Burk and Hanes were used to determine kinetic parameters of the dietary supplement inhibition. The kinetics of enzyme inhibition was assessed using the immobilized form of the inhibitor that resulted in the enzyme activity decrease at zero variations Km at the decreasing V max values which makes it possible to classify the inhibition to the linear uncompetitive type (catalyzed inhibition).
Ключевые слова
pancreatic amylase inhibitor, hydrolysis kinetics, kinetic parameters of pancreatic amylase inhibition
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Как цитировать?
Dzyuba N.A. and Prokopovich A.S. Investigation of kinetic parameters of the dietary supplement "Amil-Ing". Foods and Raw Materials, 2016, vol. 4, no. 2, pp. 128–135. DOI: 10.21179/2308-4057-2016-2-128-135.
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