Formaldehyde dehydrogenase from Pseudomonas putida: a zinc metalloenzyme.
The NAD+-dependent formaldehyde dehydrogenase from Pseudomonas putida C-83 was found to contain 4 gram atoms of zinc per mol, corresponding to 2 gram atoms of zinc per subunit monomer. Treatment of the enzyme with o-phenanthroline resulted in removal of 1 gram atom of zinc per subunit and caused a complete inactivation of the enzyme. The activity lost was restored by the addition of zinc ions, by which the zinc content was also reversed to almost the same level as that of the native enzyme. Another zinc atom that was resistant to metal chelator-treatment was liberated from the enzyme only after the irreversible denaturation of the enzyme. These results indicate that the formaldehyde dehydrogenase of P. putida is a zinc metalloenzyme and one of two zinc atoms per subunit participates in the catalytic activity of the enzyme, another zinc being presumably involved in maintaining the native conformation of the enzyme. Treatment of the enzyme with bipyridine also caused a reversible inactivation of the enzyme, but the zinc content remained unchanged. The spectrophotometric analysis indicated that the formation of a enzyme-Zn-bipyridine complex took place. Incubation of the enzyme with p-chloromercuribenzoate also resulted in a complete loss of the activity. These results suggest that an intrinsic zinc and sulfhydryl group together with NAD+ participate in the dehydrogenation reaction of substrate by the enzyme.
D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001051:Apoenzymes / Q000378:metabolism; D002384:Catalysis; D002614:Chelating Agents / Q000494:pharmacology; D055598:Chemical Phenomena; D002621:Chemistry; D004793:Enzyme Reactivators; D008670:Metals / Q000494:pharmacology; D011549:Pseudomonas / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000032:analysis; D015032:Zinc / Q000032:analysis*
D001051:Apoenzymes; D002614:Chelating Agents; D004793:Enzyme Reactivators; D008670:Metals; D013438:Sulfhydryl Compounds; C020145:formaldehyde dehydrogenase (glutathione); D000445:Aldehyde Oxidoreductases; D015032:Zinc