Identification of complexforming amylase – glutin by infrared spectroscopy

Abstract

Background. Significant shortcomings of direct enzymes input in humans restrict their availability: temperature sensitivity, pH changes and ionic strength. It stipulates perspective of studying multifunctional additives obtained by enzyme immobilization on protein matrices. The enzyme and matrix are substances of protein nature. As a result of immobilization they form strong electrostatic bonds and bonds that can be determined by infrared spectroscopy.
The article aim is to explore the possibility of formation of α-amylase – glutin complex using infrared spectroscopy.
Material and methods. The infrared absorption spectrum of samples, which were previously dried to constant mass, obtained in the discs of potassium bromide R (2 mg of substance in 200 mg of potassium bromide R), conducted in 4000 cm-1 and 400 cm-1 on infrared spectrophotometer FTIR-8400S of Shimadzu company.
Results. The possibility of using infrared spectroscopy to analyze the bond in the production of immobilized enzyme forms was showed. IR-spectrums of glutin, native and glutin-immobilized amylase were considered. Analysis of bonds in glutin molecules and all forms of immobilized enzyme was made.
Functional bonds, which provide the amylase immobilization on glutin, were determined. It is shown, that –OH, –C=O, –NH play the most important role in the formation of amylase – glutin complex.
Conclusion. Data of analyzed IR-spectrums shows a more complex molecule structure when amylase is included in the gel compared to mechanical immobilization. It confirms the hypothesis that the functional groups of glutin can form bonds with enzymes. It provides recommendation of glutin using as a matrix for the immobilization of enzymes to develop dietary supplements with directive effect.