Влияние низких температур хранения на белки инкапсулированные в альгинатные микросферы

Abstract

Вивчено вплив низьких температур зберігання на ферментативну активність білків з гемолізату, інкапсульованих у альгінатні мікросфери. Показано що при зберіганні мікросфер з інкапсульованим матеріалом протягом 18 місяців за температури -20˚С, вбудовані білки зберігають каталазну активність.

Изучено влияние низких температур хранения на ферментативную активность белков из гемо- лизата, инкапсулированных в альгинатные микросферы. Показано что при хранении микросфер с инкапсулированным материалом в течении 18 месяцев при -20˚С, встроенные белки сохраняют каталазную активность.

The protein encapsulation in matrices based on sodium alginate has already demonstrated its effectiveness in biotechnology as a stabilizing structure for encapsulated compounds. This property of alginate microspheres has been studied at room and positive temperatures. However, the stabilization of macromolecular structure is of special interest for cryobiology, since the low temperatures cause the changes in their structural and functional properties. Therefore, the study of low-temperature storage effect on encapsulated proteins in alginate microspheres provides widening of the application field for matrices based on this biopolymer. The aim of the study was to evaluate the effect of low-temperature storage of proteins from hemolysate encapsulated in alginate microspheres on their catalase activity. Object and methods. Alginate microspheres were obtained by dropwise extrusion. For this purpose sodium alginate with hemolysate was dropped into gelling solution. Solution of calcium chloride 2% served as gelling solution. Hemolysate was prepared from erythrocytes obtained from whole blood donation using ultrasound. The catalase activity was determined spectrophotometrically. In this regard the concentration of undecomposed hydrogen peroxide after its incubation with microspheres was assessed recording the colored product of hydrogen peroxide with ammonium molybdate. The size and shape of alginate microspheres were evaluated with light microscopy using MBS-10 microscope. Results. The conducted study has determined the main parameters affecting the microspheres loading from sodium alginate. They included the value of hydrogen index of alginate solution with encapsulated substance and concentration of encapsulated substance. Conclusions. It was determined that low-temperature storage of alginate microspheres with encapsulated proteins from hemolysate retained their catalase activity during 18 months at temperature -20°C.