Large-scale purification of halophilic enzymes by salting-out mediated chromatography

• Published in: Analytical biochemistry Vol. 114; no. 1; pp. 186 - 192
• Main Authors: Leicht, W., Pundak, S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.06.1981
• Subjects: Chromatography, Gel - methods; glutamate dehydrogenase; Glutamate Dehydrogenase - isolation & purification; Glutamate Dehydrogenase - metabolism; Halobacterium - enzymology; halophilic bacteria; malate dehydrogenase; Malate Dehydrogenase - isolation & purification; Malate Dehydrogenase - metabolism; Osmolar Concentration; purification; Sepharose; Temperature
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(81)90472-3

Large-scale purification of malate and glutamate dehydrogenases from extremely halophilic bacteria of the Dead Sea based on salting-out mediated chromatography techniques is described. Yields are in the range of hundreds of milligrams. Ammonium sulfate-mediated fractionation of halophilic enzymes on Sepharose is not affected by sodium chloride. The adsorption seems to weaken with increased temperatures. Higher sodium chloride concentrations, in the presence of ammonium sulfate, are necessary to elute halophilic enzymes from DEAE-cellulose as the sulfate concentration is increased. Possible applications of the saltingout mediated chromatography are discussed.