Development of methodological approaches to the purification of target proteins from marine hydrobionts of Antarctic region

M. Abramova, N. Raksha
Taras Shevchenko National University of Kyiv, Kyiv; Taras Shevchenko National University of Kyiv, Kyiv

Abstract


Enzymes from organisms that are adapted to the existence at low temperatures attract significant attention of scientists as a perspective objects not only on a practical point of view, but also as a valuable tools for conducting basic research. This is due to unusual environmental conditions (low temperature, high hydrostatic pressure, low illumination), as well as a significant level of economic profitability due to the widespread of marine hydrobionts and high efficiency of psychrophilic enzymes in comparison with their mesophilic and thermophilic analogues. The expediency of using the hydrobionts of the Antarctic region Parborlasia corrugatus and Sterechinus neumayeri as a source for producing individual enzymes was indicated by the results of electrophoretic analysis of enzymes from hydrobionts tissues extract, so it was concluded that the total extract contained a number of enzymes that were active with gelatin and fibrinogen. As a result of a combination of several chromatographic stages, which included affinity chromatography on Blue Sepharose 6 FF column and size exclusion chromatography on Superdex 75 PG and Superdex 200 PG columns, from the total hydrobiont tissue extract were obtained fractions of hydrolytic enzymes. From the total tissues extract of both hydrobionts
which was explored were isolated 4 fractions which showed gelatinase activity. Also from the tissues of Sterechinus neumayeri a fraction containing high-molecular enzymes capable of cleaving fibrinogen was isolated. The developed method of two-stage chromatography can be used further as a basis for optimizing the obtaining of enzymes of a similar spectrum of action from other objects.

Keywords


hydrobionts of Antarctic region, chromatography, electrophoresis

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Received: 06.05.2019

Revised: 06.06.2019

Accepted & Published: 06.06.2019




DOI: http://dx.doi.org/10.17721/1728_2748.2019.78.7-13

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