Abstract
Previous reports that whole cells of Methylophilus methylotrophus oxidase exogenous NADH have been investigated. Essentially identical rates of oxygen consumption were observed following the addition of methanol or NADH to whole cells. Both activities were inhibited by EDTA and hydroxylamine, but not by HQNO, and exhibited similar pH optima. Analyses of the reaction stoichiometry with NADH as substrate showed that the expected amount of oxygen was consumed, but also revealed acidification (instead of alkalinisation) and no oxidation of NADH. Further studies showed that commerical NADH is contaminated with ethanol which is oxidised to acetic acid by the low specificity methanol oxidase system present in this organism. The oxidation of exogenous NADH by whole cells of M. methylotrophus reported previously is therefore spurious.
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Abbreviations
- EDTA =:
-
ethylenediaminetetracetate
- HQNO =:
-
2−n-heptyl-4-hydroxy-quinoline-N-oxide
- DEAE =:
-
diethylaminoethyl
References
Anthony, C. (1982) The Biochemistry of methylotrophs. Academic Press, London and New York
Anthony, C. & Zatman, L. J. (1964) The microbiol oxidation of methanol: Isolation and properties of Pseudomonas sp. M27. Biochem. J. 92: 609–614
Anthony, C. & Zatman, L. J. (1965) The alcohol dehydrogenase of Pseudomonas sp. M27. Biochem. J. 96: 808–812
Beardmore-Gray, M. & Anthony, C. (1984) Methanol: Cytochrome c oxidoreductase activity of methylotrophs. In: R. L.Crawford & R. S.Hanson (eds), Microbial Growth on C1 Compounds, pp. 97–105. American Society of Microbiology, Washington.
Carver, M. A., Humphrey, K. M., Patchett, R. A. & jones, C. W. (1984) The effect of EDTA and related chelating agents on the oxidation of methanol by the methylotrophic bacterium Methylophilus methylotrophus Eur. J. Bioch. 138: 611–615
Cross, A. R. & Anthony, C. (1980) The electron transport chains of the obligate methylotroph Methylophilus methylotrophus Biochem. J. 192: 429–439
Dawson, M. J. (1982) Energy conservation in Methylophilus methylotrophus Ph. D. Thesis, University of Leicester.
Dawson, M. J. & Jones, C. W. (1981) Respiration-linked proton translocation in the obligate methylotroph Methylophilus methylotrophus Biochem. J. 194: 915–924
Duine, J. A., Frank, J. & DeRuiter, L. G. (1979) Isolation of a methanol dehydrogenase with a functional coupling to cytochrome c. J. Gen. Microbiol. 115: 523–526
Duine, J. A., Frank, J. & Westerling, J. (1978) Purification and properties of methanol dehydrogenase from Hyphomicrobium X. Biochem. Biophys. Acta 524: 277–287
Jones, C. W., Carver, M. A., Dawson, M. J. & Patchett, R. A. (1984) Respiration and ATP synthesis in Methylophilus methylotrophus. In: R. L.Crawford & R. S.Hanson (eds), Microbial Growth on C1 Compounds, pp. 134–140. American Society of Microbiology, Washington.
Sperl, G. T., Forrest, H. S. & Gibson, D. T. (1974) Substrate specificity of the purified primary alcohol dehydrogenase from methanol-oxidising bacteria. J. Bacterial 118: 541–550
White, D. C. & Sinclair, P. R. (1971) Branched electron transport systems in bacteria. Adv. Microbial. Physiol. 5: 173–211
White, D. C. & Smith, L. (1962) Haematin enzymes of Haemophilus parainfluenzae J. Biol. Chem. 237: 1332–1336
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Patchett, R.A., Jones, C.W. The apparent oxidation of NADH by whole cells of the methylotrophic bacterium Methylophilus methylotrophus . Antonie van Leeuwenhoek 52, 387–392 (1986). https://doi.org/10.1007/BF00393466
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DOI: https://doi.org/10.1007/BF00393466