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Volume 117, Issue 2

Partial Purification and Characterization of a Dye-linked Formaldehyde Dehydrogenase from X Free

Abstract

All organisms surveyed among a range of methylotrophic bacteria growing on both C and multi-carbon substrates contained a dye-linked formaldehyde dehydrogenase. The activity of this enzyme was very low in bacteria grown on C compounds when compared to the activities of other C-specific enzymes and was not induced during growth on C compounds. The enzyme was partially purified (11-fold) from methanol- and ethanol-grown X. Though not homogeneous, it did not contain methanol dehydrogenase, formate dehydrogenase or cytochrome Methylphenazonium methyl-sulphate (PMS), cytochrome , Wurster’s Blue, dichlorophenol-indophenol (DCPIP) and methylphenazonium ethosulphate (PES) could act as the primary electron acceptors in the assay system with apparent values for the first three substances of 0·69, 128 and 3·07 μ, respectively. The activity of the partially purified enzyme could be maintained over 3 months at -20° in the presence of ethanol (10%, v/v). The molecular weight of the enzyme was 83500 ± 2500. It was active from pH 6·5 to 9·0 with maximum activity at pH 7·2 using cytochrome as the electron acceptor and at pH 7·6 when PMS and DCPIP were used. Several different aldehydes could act as substrates; the apparent values for formaldehyde, acetaldehyde, propionaldehyde and heptaldehyde were 2860, 270, 13 and 2·5μ, respectively. Methanol and ethanol were not substrates. The dye-linked formaldehyde dehydrogenase is probably a general aldehyde dehydrogenase not directly involved in the dissimilation of C compounds.

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© Society for General Microbiology 1980
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1980-04-01
2024-12-03
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References

  1. Andrews P. 1965; The gel-filtration behaviour of proteins related to their molecular weights over a wide range.. Biochemical Journal 96:595–606
     [Google Scholar]
  2. Anthony C., Zatman L. J. 1964; The microbial oxidation of methanol. 2. The methanol-oxidizing enzyme of Pseudomonas sp. M27.. Biochemical Journal 92:614–621
     [Google Scholar]
  3. Anthony C., Zatman L. J. 1967; The microbial oxidation of methanol. Purification and properties of the alcohol dehydrogenase of Pseudomonas sp. M27.. Biochemical Journal 104:953–959
     [Google Scholar]
  4. Armstrong J. McD. 1964; The molar extinction coefficient of 2,6-dichlorophenol-indophenol.. Biochimica et biophysica acta 86:191–194
     [Google Scholar]
  5. Bamforth C. W., O’Connor M. L. 1979; The isolation of pleiotropic mutants of Pseudomonas aminovorans deficient in the ability to grow on methylamine and an examination of their enzymic constitution.. Journal of General Microbiology 110:143–149
     [Google Scholar]
  6. Boulton C. A., Large P. J. 1977; Synthesis of certain assimilatory and dissimilatory enzymes during bacterial adaptation to growth on tri- methylamine.. Journal of General Microbiology 101:151–156
     [Google Scholar]
  7. Colby J., Zatman L. J. 1973; Trimethylamine metabolism in obligate and facultative methylo- trophs.. Biochemical Journal 132:101–112
     [Google Scholar]
  8. Colby J., Zatman L. J. 1975; Enzymological aspects of the pathways for trimethylamine oxidation and C1 assimilation in obligate methylo- trophs and restricted facultative methylotrophs.. Biochemical Journal 148:513–520
     [Google Scholar]
  9. Eisenthal R., Cornish-Bowden A. 1974; The direct linear plot. A new graphical procedure for estimating enzyme kinetic parameters.. Biochemical Journal 139:715–720
     [Google Scholar]
  10. Harder W., Attwood M. M. 1978; Biology, physiology and biochemistry of hyphomicrobia.. Advances in Microbial Physiology 17:303–359
     [Google Scholar]
  11. Harder W., Attwood M. M., Quayle J. R. 1973; Methanol assimilation by Hyphomicrobium sp.. Journal of General Microbiology 78:155–163
     [Google Scholar]
  12. Heptinstall J., Quayle J. R. 1970; Pathways leading to and from serine during growth of Pseudomonas AMI on C1 compounds of succinate.. Biochemical Journal 117:563–572
     [Google Scholar]
  13. Johnson P. A., Quayle J. R. 1964; Microbial growth on C1 compounds. 6. Oxidation of methanol, formaldehyde and formate by methanol- grown Pseudomonas AM1.. Biochemical Journal 93:281–290
     [Google Scholar]
  14. Maitzel J. V. 1971; Polyacrylamide gel electrophoresis of viral proteins.. Methods in Virology 5:179–246
     [Google Scholar]
  15. Mehta R. J. 1973; Studies on methanol-oxidizing bacteria. II. Purification and properties of methanol dehydrogenase from Pseudomonas RJ1.. Antonie van Leeuwenhoek 39:303–312
     [Google Scholar]
  16. Mehta R. J. 1975; A novel inducible formaldehyde dehydrogenase of Pseudomonas sp. RJ1.. Antonie van Leeuwenhoek 41:89–95
     [Google Scholar]
  17. Patel R. N., Hoare D. S. 1971; Physiological studies of methane and methanol-oxidizing bacteria: oxidation of C1 compounds by Methylococcus capsulatus.. Journal of Bacteriology 107:187–192
     [Google Scholar]
  18. Patel R. N., Bose H. R., Mandy W. J., Hoare D. S. 1972; Physiological studies of methane and methanol-oxidizing bacteria: comparison of a primary alcohol dehydrogenase from Methylococcus (Texas strain) and Pseudomonas sp. M27.. Journal of Bacteriology 110:570–577
     [Google Scholar]
  19. Stirling D. I., Dalton H. 1978; Purification and properties of an NAD(P)+-linked formaldehyde dehydrogenase from Methylococcus capsulatus (Bath).. Journal of General Microbiology 107:19–29
     [Google Scholar]
  20. Strøm T., Ferenci T., Quayle J. R. 1974; The carbon assimilation pathways of Methylococcus capsulatus, Pseudomonas methanica and Methylo-sinus trichosporium (OB3B) during growth on methane.. Biochemical Journal 144:465–476
     [Google Scholar]
  21. Van Dijken J. P., Otto R., Harder W. 1975; Oxidation of methanol formaldehyde and formate by catalase purified from methanol-grown Hansenula polymorpha.. Archives of Microbiology 106:221–226
     [Google Scholar]
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Partial Purification and Characterization of a Dye-linked Formaldehyde Dehydrogenase from Hyphomierobium X
Microbiology 117, 305 (1980); https://doi.org/10.1099/00221287-117-2-305
/content/journal/micro/10.1099/00221287-117-2-305
/content/journal/micro/10.1099/00221287-117-2-305
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