Collect. Czech. Chem. Commun. 1998, 63, 851-856

Single-Step Separation of Lactate Dehydrogenase Using Thiophilic Chromatography

Milena Kmínková and Jiří Kučera

Food Research Institute, Radiová 7, CZ-102 31 Prague 10, Czech Republic


Lactate dehydrogenase (EC from carp (Cyprinus carpio) hepatopancreas was purified by the single-step chromatography on thiophilic sorbent. Hydrophobic negative sorption was used as negative adsorption step for final purification. Final specific activity was 8.88 U/mg protein and the yield 84%. The enzyme is not adsorbed during hydrophobic interaction chromatography, but proteolytic activity is adsorbed completely on hydrophobic sorbent. Thus hydrophobic adsorption can be used as a prepurification step.

Keywords: Lactate dehydrogenase; Carp; Thiophilic chromatography; Enzymes; Oxidoreductases; Cyprinus carpio.