1887

Abstract

Both the soluble cytochromes of the obligate methylotroph were rapidly autoreducible at high pH. The intramolecular autoreduction mechanism was also involved in the reduction of the cytochrome by methanol dehydrogenase which occurred in the absence of methanol. Pure soluble methanol dehydrogenase was shown to be able to catalyse the methanol-dependent reduction of pure cytochrome from and from the facultative methylotroph Pseudomonas AM1 by coupling oxidation of the bacterial cytochrome to the reduction of a large excess of mammalian cytochrome . Only one of the two cytochromes (cytochrome of each organism) could react with methanol dehydrogenase to give methanol:cytochrome oxidoreductase activity. This activity, using proteins from , was independent of pH between pH 7·0 and 9·0 and ammonia was not required. By contrast, the pH optimum for the system from Pseudomonas AM1 was 9.0 and activity was stimulated about fourfold by NHCl. The product of methanol oxidation was formaldehyde, which was also a substrate for the oxidoreductase system. During formaldehyde oxidation two molecules of cytochrome were reduced for every molecule of formaldehyde oxidized. In a survey of methanol dehydrogenases and cytochromes from Pseudomonas AM1, and the facultative autotroph , it was shown that, of the two soluble cytochromes found in each methylotroph only one was able to react with methanol dehydrogenase. The cytochrome from and the cytochrome (2) of were specific, only reacting with methanol dehydrogenase from the same organism, whereas the cytochrome of AM1 reacted with all three methanol dehydrogenases tested.

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1983-04-01
2021-07-24
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References

  1. Alefounder P. R., Ferguson S. J. 1981; A periplasmic location for methanol dehydrogenase from Paracoccus denitrificans. Implications for proton pumping by cytochrome aa3. Biochemical and Biophysical Research Communications 98:778–784
    [Google Scholar]
  2. Anthony C. 1975; The microbial oxidation of Q compounds. The cytochromes of Pseudomonas AM 1. Biochemical Journal 146:289–298
    [Google Scholar]
  3. Anthony C. 1981; Electron transport in methylotrophic bacteria. In Microbial Growth on C, Compounds pp. 220–230 Edited by Dalton H. London: Heyden;
    [Google Scholar]
  4. Anthony C. 1982a The Biochemistry of Methylotrophs pp. 167–187 London: Academic Press;
    [Google Scholar]
  5. Anthony C. 1982b The Biochemistry of Methylotrophs pp. 219–244 London: Academic Press;
    [Google Scholar]
  6. Anthony C., Zatman L. J. 1964; The microbial oxidation of methanol.The methanol-oxidising enzyme of Pseudomonas sp. M27. Biochemical Journal 92:614–621
    [Google Scholar]
  7. Anthony C., Zatman L. J. 1965; The microbial oxidation of methanol.The alcohol dehydrogenase of Pseudomonas sp. M27. Biochemical Journal 96:808–812
    [Google Scholar]
  8. Bamforth C. W., Quayle J. R. 1978; Aerobic and anaerobic growth of Paracoccus denitrificans on methanol. Archives of Microbiology 119:91–97
    [Google Scholar]
  9. Beardmore-Gray M. 1982 Cytochrome c: its role in methylotrophic bacteria. Ph.D. thesis, Southampton University
    [Google Scholar]
  10. Beardmore-Gray M., O’Keeffe D. T., Anthony C. 1982; The autoreducible cytochromes c of the methylotrophs, Methylophilus methylotrophus and Pseudomonas AM1. Biochemical Journal 207:161–165
    [Google Scholar]
  11. Capaldi R. A., Hayashi H. 1972; The polypeptide composition of cytochrome oxidase from beef-heart mitochondria. FEBS Letters 26:261–263
    [Google Scholar]
  12. Cross A. R., Anthony C. 1980a; The purification and properties of the soluble cytochromes cof the obligate methylotroph, Methylophilus methylotrophus. Biochemical Journal 192:421–427
    [Google Scholar]
  13. Cross A. R., Anthony C. 1980b; The electron transport chains of the obligate methylotroph, Methylophilus methylotrophus. Biochemical Journal 192:429–439
    [Google Scholar]
  14. Duine J. A., Frank J. 1981; Methanol dehydrogenase: a quinoprotein. In Microbial Growth on C, Compounds pp. 31–41 Edited by Dalton H. London: Heyden;
    [Google Scholar]
  15. Duine J. A., Frank J., De Ruiter L. G. 1979; Isolation of a methanol dehydrogenase with a functional coupling to cytochrome c. Journal of General Microbiology 115:523–526
    [Google Scholar]
  16. Dunstan P. M., Anthony C., Drabble W. T. 1972; Microbial metabolism of C1 and C2 compounds : the involvement of glycollate in the metabolism of ethanol and of acetate by Pseudomonas AML. Biochemical Journal 128:99–106
    [Google Scholar]
  17. Ghosh R. 1980 Studies on the methanol dehydrogenase from Methylophilus methylotrophus. Ph.D. thesis, Sheffield University
    [Google Scholar]
  18. Ghosh R., Quayle J. R. 1981; Purification and properties of the methanol dehydrogenase from Methylophilus methylotrophus. Biochemical Journal 199:245–250
    [Google Scholar]
  19. Goldberg I. 1976; Purification and properties of a methanol-oxidising enzyme in PseudomonasC. European Journal of Biochemistry 63:233–240
    [Google Scholar]
  20. Jones C. W., Kingsbury S. A., Dawson M. J. 1982; The partial resolution and dye-mediated reconstitution of methanol-oxidase activity in Methylophilus methylotrophus. FEMS Microbiology Letters 13:195–200
    [Google Scholar]
  21. Nash T. 1953; The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochemical Journal 55:416–421
    [Google Scholar]
  22. Ohta S., Tobari J. 1981; Two cytochromes c of Methylomonas J. Journal of Biochemistry 90:215–224
    [Google Scholar]
  23. O’Keeffe D. T., Anthony C. 1980a; The two cytochromes c in the facultative methylotroph, Pseudomonas AM1. Biochemical Journal 192:411–419
    [Google Scholar]
  24. O’Keeffe D. T., Anthony C. 1980b; The interaction between methanol dehydrogenase and the autoreducible cytochromes c of the facultative methylotroph, Pseudomonas AM1. Biochemical Journal 190:481–484
    [Google Scholar]
  25. Sperl G. T., Forrest H. S., Gibson D. T. 1974; Substrate specificity of the purified primary alcohol dehydrogenases from methanol-oxidising bacteria. Journal of Bacteriology 118:541–550
    [Google Scholar]
  26. Tanaka K., Takahashi M., Asada K. 1978; Isolation of monomeric cytochrome/from Japanese radish and a mechanism of autoreduction. Journal of Biological Chemistry 253:7397–7403
    [Google Scholar]
  27. Van Verseveld H. W., Stouthamer A. H. 1978; Electron transport chain and coupled oxidative phosphorylation in methanol grown Paracoccus denitrificans. Archives of Microbiology 118:13–20
    [Google Scholar]
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