Affinity chromatography of bacterial lactate dehydrogenases.
The affinity system used was the immobilized oxamate derivative previously used to purify mammalian lactate dehydrogenases. The bacterial dehydrogenases specific for the L-stereoisomer of lactate behaved in the same way as the mammalian enzymes, binding strongly in the presence of NADH. The D-lactate-specific enzymes, however, did not show any biospecific affinity for this gel. The L-specific enzymes could be purified to homogeneity in one affinity-chromatographic step. The D-specific enzymes could be efficiently separated from the L-specific ones and could then be further purified on an immobilized NAD derivative. The mechanism of activation of the lactate dehydrogenase from Streptococcus faecalis by fructose 1,6-bisphosphate was investigated by using the immobilized oxamate gel.
D001419:Bacteria / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods; D005635:Fructosediphosphates / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D010072:Oxamic Acid; D013237:Stereoisomerism
D005635:Fructosediphosphates; D007770:L-Lactate Dehydrogenase; D010072:Oxamic Acid