Purification and properties of lactate dehydrogenase from Nocardia asteroides.
The lactate dehydrogenase (LDH) from Nocardia asteroides was purified to homogeneity by ammonium sulphate precipitation, gel filtration on Sephadex G-150 and DEAE-Sepharose column chromatography. The purified enzyme showed a single band in native condition which indicated its homogeneity. SDS-PAGE of the purified enzyme showed the presence of three bands which correspond to molecular weights of 60, 66 and 74 kDa. The pH and temperature optima of the purified enzyme were 9.5 and 50 degrees C, respectively. The metal ions Mn++, Fe++, Co++, Mg++ and Ca++, increased the purified LDH activity. On the other hand, enzyme activity was completely inhibited by CuCl2. Potassium chloride, ammonium sulphate and sodium chloride did not alter the enzyme activity. The purified enzyme exhibited a Km value of 1.6 x 10(-5) M for pyruvate.
Ike
J
J
Department of Biology, Fisk University, Nashville, Tennessee 37208.
Sangan
P
P
Gunasekaran
M
M
D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003300:Copper / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D009616:Nocardia asteroides / Q000201:enzymology*; D013696:Temperature
D008670:Metals; D003300:Copper; C028419:cuprous chloride; D007770:L-Lactate Dehydrogenase