Ladungskontrollierte hydrophobe chromatographie: Ein effektives trennprinzip für proteine

More than twenty enzymes (oxidoreductases, hydrolases, lyases, transferases) from crude bacterial extracts or commercial preparations have been purified by charge-controlled hydrophobic chromatography on 10-carboxydecyl-Sepharose. The enzymes were adsorbed on columns in the presence of high concentrations of structure-forming anions (phosphate, sulphate, citrate) and were eluted by decreasing the concentration of the salts. As a rule crystalline enzyme preparations were obtained by dialysis of eluates against ammonium sulphate solutions. These results suggest the existence of a true affinity principle based on the ability of structure-forming anions to modify the enzyme conformation. The only supposition is the immobilization of a ligand with a specific combination of hydrophobic and ionic properties in that manner, that the columns are not able to bind enzymes in the presence of dilute buffer solutions.