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Studies of the Processes of the Trypsin Interactions with Ion Exchange Fibers and Chitosan

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Abstract

A localization of charged and hydrophobic amino acid residues in the trypsin molecule was studied, and a percentage of the amino acids of different types on a surface of the enzyme globule was determined. The charged and hydrophobic amino acid residues were shown to be irregularly distributed on the protein surface and to form local clusters. The VION KN-1 and VION AN-1 fibers and chitosan were found to be promising carriers for the trypsin immobilization, because an adsorption on these fibers provided the preservation of 54, 58 and 65% of the catalytic activity of the native enzyme in solution, respectively (measured according to the hydrolysis rate of the bovine serum albumin). The IR spectra of the native (free) enzyme and the enzyme immobilized on the polymeric supports were analyzed. Electrostatic interactions and hydrogen bonds were shown to be dominant during the trypsin adsorption on the VION fibers. Carboxyl groups of the VION KN-1 interacted with positively charged regions of the molecule which contained His, Lys, and Arg. A large number of amino groups of the VION AN-1 and chitosan created an excessive positive charge which, possibly, provided a binding to the negatively charged Asp and Glu. However, hydrophobic interactions in which Gly, Ala, Tyr, Val, Phe, Pro, and Leu were involved became the most important for the trypsin adsorption on chitosan.

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ACKNOWLEDGMENTS

The experiments were performed using the scientific and technical base of the Center of the Collective Use of the scientific equipment of the Voronezh State University.

Funding

This study was supported by the Grant of the President of the Russian Federation for the state support of the young Russian DPhil scientists MD-1982.2020.4, project 075-15-2020-325.

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Authors and Affiliations

  1. Voronezh State University, 394018, Voronezh, Russia

    S. M. Pankova, F. A. Sakibaev, M. G. Holyavka, A. N. Lukin & V. G. Artyukhov

  2. Moscow Institute of Physics and Technology, 141701, Dolgoprudnyi, Moscow oblast, Russia

    Y. M. Vyshkvorkina

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  1. S. M. Pankova
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  2. F. A. Sakibaev
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  3. M. G. Holyavka
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Correspondence to M. G. Holyavka.

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Translated by L. Onoprienko

Abbreviations: BSA, bovine serum albumin.

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Pankova, S.M., Sakibaev, F.A., Holyavka, M.G. et al. Studies of the Processes of the Trypsin Interactions with Ion Exchange Fibers and Chitosan. Russ J Bioorg Chem 47, 765–776 (2021). https://doi.org/10.1134/S1068162021030146

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