Competitive elution of lactate dehydrogenase from Cibacron Blue—bead cellulose with Cibacron Blue—dextrans
1990; Elsevier BV; Volume: 510; Linguagem: Inglês
10.1016/s0021-9673(01)93753-7
ISSN1873-3778
AutoresDanica Mislovičová, Peter Gemeiner, Eva Stratilová, Marta Horváthova,
Tópico(s)Enzyme Structure and Function
ResumoAbstract The efficiencies of elution of lactate dehydrogenase (LDH) from Cibacron Blue (CB)—bead cellulose with eluents ensuring competitive (Cibacron Blue—dextran), biomimetic NADH) and displacing (KCl) mechanisms were compared. Competitive elution with CB—dextran T 10 was shown to be the most effective providing a 38 fold purified enzyme in 83% yield. As shown by fast protein liquid chromatography and polyacrylamide gel electrophoresis, this LDH preparation was free from protein contaminants but contained CB—dextran. CB—dextran was then removed by ion-exchange chromatography and the yield of LDH decreased to 62%. When using a longer column, the enzyme was resolved partially in two fractions. The isoelectric point of the main fraction was 7.3.
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