Novel antioxidant role of alcohol dehydrogenase E from Escherichia coli.
Alcohol dehydrogenase E (AdhE) is an Fe-enzyme that, under anaerobic conditions, is involved in dissimilation of glucose. The enzyme is also present under aerobic conditions, its amount is about one-third and its activity is only one-tenth of the values observed under anaerobic conditions. Nevertheless, its function in the presence of oxygen remained ignored. The data presented in this paper led us to propose that the enzyme has a protective role against oxidative stress. Our results indicated that cells deleted in adhE gene could not grow aerobically in minimal media, were extremely sensitive to oxidative stress and showed division defects. In addition, compared with wild type, mutant cells displayed increased levels of internal peroxides (even higher than those found in a Delta katG strain) and increased protein carbonyl content. This pleiotropic phenotype disappeared when the adhE gene was reintroduced into the defective strain. The purified enzyme was highly reactive with hydrogen peroxide (with a Ki of 5 microM), causing inactivation due to a metal-catalyzed oxidation reaction. It is possible to prevent this reactivity to hydrogen peroxide by zinc, which can replace the iron atom at the catalytic site of AdhE. This can also be achieved by addition of ZnSO4 to cell cultures. In such conditions, addition of hydrogen peroxide resulted in reduced cell viability compared with that obtained without the Zn treatment. We therefore propose that AdhE acts as a H2O2 scavenger in Escherichia coli cells grown under aerobic conditions.
Echave
Pedro
P
Departament de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, 25198 Lleida, Spain.
Tamarit
Jordi
J
Cabiscol
Elisa
E
Ros
Joaquim
J
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000502:physiology*; D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000502:physiology*; D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D020134:Catalytic Domain; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D029968:Escherichia coli Proteins; D017353:Gene Deletion; D005947:Glucose / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000737:chemistry; D007700:Kinetics; D009097:Multienzyme Complexes / Q000737:chemistry / Q000502:physiology*; D009154:Mutation; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D010641:Phenotype; D010802:Phylogeny; D013997:Time Factors; D015032:Zinc / Q000737:chemistry
D000975:Antioxidants; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D016877:Oxidants; D006861:Hydrogen Peroxide; D000426:Alcohol Dehydrogenase; C068383:adhE protein, E coli; D000445:Aldehyde Oxidoreductases; D005947:Glucose; D015032:Zinc; D010100:Oxygen