Simple efficient methods for the isolation of malate dehydrogenase from thermophilic and mesophilic bacteria.
Malate dehydrogenase from a number of bacteria drawn from several genera and representing the mesophilic, moderately thermophilic and extremely thermophilic classes was isolated by procedures which involve only a small number of steps (in most cases only two), of which the key one is affinity chromatography on 5'-AMP--Sepharose and/or on NAD+--hexane--agarose. Electrophoretic analysis of the native enzymes in polyacrylamide gel and of the denaturated enzymes in sodium dodecyl sulphate/polyacrylamide gel revealed no significant protein impurity in the purified preparations. The yields ranged from about 40% to over 80%. The malate dehydrogenases from the extreme thermophiles and from some of the moderate thermophiles are appreciably less efficient catalytically than their mesophilic homologues.
D000249:Adenosine Monophosphate; D001407:Bacillus / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D008291:Malate Dehydrogenase / Q000302:isolation & purification*; D008722:Methods; D011549:Pseudomonas / Q000201:enzymology; D013824:Thermus / Q000201:enzymology
D000249:Adenosine Monophosphate; D008291:Malate Dehydrogenase