Разделение изоформ L-лактатдегидрогеназы из галоалкалофильной бактерии Rhodovulum steppense штамма A-20s методом ионообменной хроматографии

Т. В. Сорокина; В. М. Ларченков; Х. А. Абдуллах; Ж. А. Целуйко; М. И. Фалалеева; А. Т. Епринцев

doi:10.17308/sorpchrom.2015.15/325

Т. В. Сорокина Sorokina Tatyana V. – the student of Voronezh State University, Voronezh
В. М. Ларченков Larchenkov Vladimir M. – the postgraduate student, department of biochemistry and physiology of cell, Voronezh State University, Voronezh, email: larchenkoff.vladimir@yandex.ru
Х. А. Абдуллах Abdullah Hasan Ahmed – the postgraduate student, department of biochemistry and physiology of cell, Voronezh State University, Voronezh
Ж. А. Целуйко Tseluyko Zhanna A. – the student of Voronezh State University, Voronezh
М. И. Фалалеева Falaleeva Marina I. – Ph. D. (biology), associate prof., department of biochemistry and physiology of cell, Voronezh State University, Voronezh
А. Т. Епринцев Eprintsev Alexander T. – prof., grand Ph. D. (biology), department of biochemistry and physiology of cell, head pf chair, Voronezh State University, Voronezh

DOI: https://doi.org/10.17308/sorpchrom.2015.15/325

Keywords: lactate dehydrogenase, step of purification, ion exchange chromatography, electrophoresis, isoenzyme, Michaelis constant

Abstract

The aim of this research work is a description of some catalytic characteristics of lactate
dehydrogenase from haloalkaliphilic bacteria Rhodovulum steppense strain A-20s and creation of methods
for extracting of this enzyme from bacterial cells. Rhodovulum sulphidophilum is cultivated on a salt medium
with the sodium acetate and yeast extract as a carbon sourses. The methods of ion exchange and gel
chromatography were used for purification of enzyme. Electrophoretic analysis is realized by Davis method,
enzyme staining is realized by tetrazolium method. The presence of two L-lactate dehydrogenase isoforms in
Rhodovulum steppense is determined. The pH optimum for the first isoform is 7.5-7.75, and Michaelis
constant value for this isoform is 0.14. Electrophoretic mobility coefficient values are 0.329 for first isoform
and 0.714 for second isoform. Our results are confirmed the presence of L-lactate dehydrogenase in the
Rhodovulum steppense in phototrophic conditions. This data may be useful for the creation of metabolism
model of purple bacteria and biotechnological application of these organisms.

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Differentiation of lactate dehydrogenase isoforms using ion exchange chromatography

Abstract

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