Skip to main content
SpringerLink
Log in

Studies on methanol — oxidizing yeast

III. Enzyme

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Oxidation of methanol, formaldehyde and formic acid was studied in cells and cell-free extract of the yeastCandida boidinii No. 11Bh. Methanol oxidase, an enzyme oxidizing methanol to formaldehyde, was formed inducibly after the addition of methanol to yeast cells. The oxidation of methanol by cell-free extract was dependent on the presence of oxygen and independent of any addition of nicotineamide nucleotides. Temperature optimum for the oxidation of methanol to formaldehyde was 35°C, pH optimum was 8.5. The Km for methanol was 0.8mM. The cell-free extract exhibited a broad substrate specificity towards primary alcohols (C1-C6). The activity of methanol oxidase was not inhibited by lnni KCN, EDTA or monoiodoacetic acid. The strongest inhibitory action was exerted byp-chloromercuribenzoate. Both the cells and the cell-free extract contained catalase which participated in the oxidation of m||ethanol to formaldehyde; the enzyme was constitutively formed by the yeast. The pH optimum for the degradation of H2O2 was in the same range as the optimum fop methanol oxidation,viz. at 8.5. Catalase was more resistant to high pH than methanol oxidase. The cell-free extract contained also GSH-dependent NAD-formaldehyde dehydrogenase with Km= 0.29mM and NAD-formate dehydrogenoae withK m = 55mM.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anthony C., Zatman L. J.: The microbial oxidation of methanol. 2. The methanol-oxidizing enzyme oPaeudomonas sp. M27.Biochem. J.92, 614 (1964).

    PubMed  CAS  Google Scholar 

  • Anthony C., Zatman L. J.: The microbial oxidation of methanol. The alcohol dehydrogenase ofPaeudomonas sp. M27.Biochem. J.96, 808 (1965).

    PubMed  CAS  Google Scholar 

  • Anthony C., Zatman L. J.: The microbial oxidation of methanol. Purification and properties of the alcohol dehydrogenase ofPseudomonas sp. M27.Biochem. J.104, 953 (1967).

    PubMed  CAS  Google Scholar 

  • Fujit T., Tonomura K.: Oxidation of methanol, formaldehyde and formate by aCandida species.Agr. Biol. Chem.36, 2297 (1972).

    Google Scholar 

  • Harrington A. A., Kallio R. E.: Oxidation of methanol and formaldehyde byPseudomonas methanica.Can. J. Microbiol.6, 1 (1960).

    PubMed  CAS  Google Scholar 

  • Hayaishi O., Katagiri M., Rothberg S.: Studies on oxygenases. Pyrocatechase.J. Biol. Chem.229, 905 (1957).

    PubMed  CAS  Google Scholar 

  • Huggett A. G., Nixon D. A.: Enzymic determination of blood glucose.Biochem. J.66, 12p (1957).

    Google Scholar 

  • Johnson P. A., Quayle J. R.: Microbial growth on C1 compounds. 6. Oxidation of methanol, formaldehyde and formate by methanol-grownPseudomonas AM1.Biochem. J.93, 281 (1964).

    PubMed  CAS  Google Scholar 

  • Kaneda T., Roxburgh J. M.: A methanol-utilizing bacterium. II. Studies on the pathway of methanol assimilation.Can. J. Microbiol.5, 187 (1959).

    Article  PubMed  CAS  Google Scholar 

  • Kaprálek F.: The physiology of ribofiavin production byEremothecium ashbyi.J. Gen. Microbiol.29, 403 (1962).

    Google Scholar 

  • Kato N., Tamaoki T., Tani Y., Ogata K.: Purification and characterization of formaldehyde dehydrogenase in.a methanol-utilizing yeast,Kloeckera sp. No. 2201.Agr. Biol. Chem.36, 2411 (1972).

    CAS  Google Scholar 

  • Lodder J.: The Yeasts. A taxonomic study. North-Holland Publ. Co. Amsterodam, 1970.

    Google Scholar 

  • Lotspeich W. D., Peters R. A.: The action of sulphydryl inhibitors upon isocitric dehydrogenase with especial reference to the behaviour of some trivalent arsenicals.Biochem. J.49, 704 (1951).

    PubMed  CAS  Google Scholar 

  • Lowry O. H., Rosebrough N. J., Farr N. J., Randall R. J.: Protein measurement with the folin phenol reagent.J. Biol. Chem.193, 265 (1951).

    PubMed  CAS  Google Scholar 

  • MacFadikn D. A.: Estimation of formaldehyde in biological mixtures.J. Biol. Chem.158, 107 (1945).

    Google Scholar 

  • Mahler H. R., Mackler B., Green D. E., Bock R. M.: Studies on metalloflavoproteins. III. Aldehyde oxidase: A molybdoflavoprotein.J. Biol. Chem.210, 465 (1954).

    PubMed  CAS  Google Scholar 

  • Mehta R. J.: Studies on methanol-oxidizing bacteria. II. Purification and properties of methanol dehydrogenase from Pseudomonas RJ1.Ant. Leeuwenhoek J. Microbiol. Ser.39, 303 (1973).

    Article  CAS  Google Scholar 

  • Ogata K., Nishikawa H., Ohsugi M.: A yeast capable of utilizing methanol.Agr. Biol. Chem.33, 1519 (1969).

    CAS  Google Scholar 

  • Patel R. N., Hoare D. S.: Physiological studies of methane and methanol-oxidizing bacteria: oxidation of C-1 compounds byMethylococeus capsulatus.J. Bacteriol.107, 187 (1971).

    PubMed  CAS  Google Scholar 

  • Quayle J. R.: The metabolism of one-carbon compounds by microorganisms.Adv. Microbiol. Physiol.7, 119 (1972).

    CAS  Google Scholar 

  • Rajagopalan K. V., Fridovich I., Handler P.: Hepatic aldehyde oxidase. I. Purification and properties.J. Biol. Chem.237, 922 (1962).

    PubMed  CAS  Google Scholar 

  • Roggenkamp R., Sahm H., Wagner F.: Microbial assimilation of methanol, induction and function of catalase inCandida boidinii.FEBS Lett.41, 283 (1974).

    Article  PubMed  CAS  Google Scholar 

  • Sahm H., Wagner F.: Microbial assimilation of methanol. The ethanoland methanol-oxidizing enzymes of the yeastCandida boidinii.Europ. J. Biochem.36, 250 (1973a).

    Article  PubMed  CAS  Google Scholar 

  • Sahm H., Wagner F.: Microbial assimilation of methanol. Properties of formaldehyde dehydrogenase and formate dehydrogenase fromCandida boidinii.Arch. Mikrobiol.90, 263 (1973b).

    Article  PubMed  CAS  Google Scholar 

  • Tani Y., Miya T., Nishikawa H., Ogata K.: The microbial metabolism of methanol. Part I. Formation and crystallization of methanol-oxidizing enzyme in methanol-utilizing yeastKloeckera sp. No. 2201.Agr. Biol. Chem.36, 68 (1972a).

    CAS  Google Scholar 

  • Tani Y., Miya T., Ogata K.: The microbial metabolism of methanol. Part II. Properties of crystalline alcohol oxidase fromKloeckera sp. No. 2201.Agr. Biol. Chem.36, 76 (1972b).

    CAS  Google Scholar 

  • Volfová O.: Oxidation of methanol by aCandida species. Proc. 4th Internat. Symp. Yeasts, Vienna, Austria 1974, Part I, A 22.

  • VoLfová O., Panos J., Pecka K.: Studies on methanol-oxidizing yeast. II. Lipids.Folia Microbiol.19, 373 (1974).

    Google Scholar 

  • Volfová O., Pilát P.: Studies on methanol-oxidizing yeast. I. Isolation and growth studies.Folia Micro biol.19, 249 (1974).

    Article  Google Scholar 

  • Volfová O., Sikyta B., Pilát P., Kalasová M.: A new production strain ofCandida boidinii no. 11Bh. Czechosl. Patent Appl. PV 3340–74, 1974.

  • Webb J. L.: Enzyme and metabolic inhibitors. Vol. II, p. 635. New York and London, Academic Press 1966.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Technical Microbiology, Institute of Microbiology, Czechoslovak Academy of Sciences, 142 20, Prague 4

    O. Volfoá

Authors
  1. O. Volfoá
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Volfoá, O. Studies on methanol — oxidizing yeast. Folia Microbiol 20, 307–319 (1975). https://doi.org/10.1007/BF02878112

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02878112

Keywords

Search

Navigation