The two alcohol dehydrogenases of Zymomonas mobilis. Purification by differential dye ligand chromatography, molecular characterisation and physiological roles.
The two alcohol dehydrogenases found in Zymomonas mobilis have each been purified using dye-ligand chromatography and affinity elution with nucleotides. The isoenzyme with lower electrophoretic mobility (ZADH-1) is a zinc enzyme with properties essentially similar to preparations described elsewhere. The faster isoenzyme (ZADH-2) accounted for some 90% of the ethanol-oxidizing activity in freshly prepared extracts and corresponded to the iron-activated enzyme previously described. This enzyme was inactivated by zinc; activity could only be retained during purification by including either ferrous ions or cobaltous ions in the buffers. ZADH-2 has relatively low acetaldehyde reductase activity; consequently ZADH-1 is responsible for about half of the physiological activity (acetaldehyde reduction) in Zymomonas cells. Kinetic studies showed that ZADH-2 is activated by ethanol in both reaction directions; a hypothesis for the mechanism of activation is presented. Metal ion analyses of ZADH-2 prepared in the presence of iron or cobalt indicated one atom of the relevant metal per subunit, with no significant zinc content. N-terminal sequence analyses showed that the ZADH-1 has some homology with the Bacillus stearothermophilus enzyme, whereas ZADH-2 resembles the yeast enzyme more closely.
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000145:classification / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D004396:Coloring Agents; D004789:Enzyme Activation / Q000187:drug effects; D006090:Gram-Negative Bacteria / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000032:analysis; D007527:Isoenzymes / Q000302:isolation & purification*; D007700:Kinetics; D008970:Molecular Weight
D004396:Coloring Agents; D007527:Isoenzymes; D007501:Iron; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase