Chromatography in presence of high concentrations of salts on columns of celluloses with and without ion exchange groups (hydrogen bond chromatography). Its application to purification of yeast enzymes.
1. All the water-soluble yeast enzymes tested, which were only partially precipitated at best in the presence of high concentration of salts such as ammonium sulfate or sodium formate, were adsorbed on a column of cellulose in the presence of the same concentrations of the salts, and the adsorbed enzymes were chromatographically eluted by decreasing the concentration of the salts. 2. Even in the presence of high concentration of the salts, the adsorbed enzymes were eluted by urea or by "hydroxy-rich" reagents such as sucrose. 3. Under the experimental conditions used, the salt concentrations required for elution of the adsorbed enzymes were lower with cellulose than with DEAE-cellulose, CM-cellulose, or P-cellulose, indicating that ion exchange groups, either cationic or anionic, affected the adsorption, although the ion exchange groups of DEAE-cellulose, CM-cellulose, and P-cellulose were weakly but definitely functional as ion exchangers even in the presence of high concentrations of the salts. 4. The principal attractive force between cellulose and the enzyme was deduced to be due to hydrogen bonding. 5. This hydrogen bond chromatography was applied for the purification of some yeast enzymes.
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification; D002845:Chromatography / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004798:Enzymes / Q000302:isolation & purification*; D005956:Glucose-6-Phosphate Isomerase / Q000302:isolation & purification; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D005980:Glutathione Reductase / Q000302:isolation & purification; D005991:Glycerol Kinase / Q000302:isolation & purification; D006593:Hexokinase / Q000302:isolation & purification; D006860:Hydrogen Bonding; D007521:Isocitrate Dehydrogenase / Q000302:isolation & purification; D008291:Malate Dehydrogenase / Q000302:isolation & purification; D011770:Pyruvate Kinase / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D004798:Enzymes; D000429:Alcohol Oxidoreductases; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D005980:Glutathione Reductase; D006593:Hexokinase; D005991:Glycerol Kinase; D011770:Pyruvate Kinase; D005956:Glucose-6-Phosphate Isomerase