Folia Microbiologica

, Volume 37, Issue 4, pp 304–310 | Cite as

Physiological and morphological acid resistance ofCandida boidinii

  • O. Volfová
  • M. Anděrová
  • Z. Žižka


The growth ofCandida boidinii strain 2 in a methanol-limited chemostat at a dilution rate of 0.1/h and a low extracellular pH (2.8–4.0) is characterized by a maximum yield coefficient referred to the methanol consumedY S of 0.4 g/g and a maximum cell content of nitrogenous compounds of 60%. The cell proteins are rich in essential amino acids. At pH<2.6 or >4.0 the cell concentration decreases due to lower growth rate, accompanied by increased metabolic quotientsQ S,Q CO2 andQ form, and increased activities of dissimilating dehydrogenases. The activity of alcohol oxidase (AO) in intact cells (0.54 IU/mg protein) was unaffected by pH 2.8–3.8 although in a cell-free extract the AO activity decreased at these low pH values after a 10-min incubation. The lower AO activity in cells at pH<2.8 and pH>3.8 brought about increased residual methanol levels in the medium, and also an increased level of riboflavin phosphate, arising probably by the release of FAD from active AO. Catalase activity was completely pH-independent. Cell morphology also showed no changes at pH 2.8–4.2, formation of cell chains being observed only at pH<2.8. However, the ultrastructure of cells grown in the chemostat at pH 2.6, however, did not evince any changes as compared with cells grown, at higher pH apart from a lag in cytokinesis. These findings, which point to acid resistance of strain 2, make it possible to produce biomass from methanol, with a high content of valuable proteins and AO, under nonsterile conditions.


Dilution Rate Alcohol Oxidase Methylotrophic Yeast Metabolic Quotient Formaldehyde Dehydrogenase 


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  1. Aminova L.R., Kyslíková E., Volfová O., Trotsenko Y.A.: Characterization of catalase-negative mutants of methylotrophic yeastHansenula polymorpha.Folia Microbiol. 36, 158–163 (1991).Google Scholar
  2. Douma A.C., Veenhuis M., Koning W., de Evers M., Harder W.: Dihydroxyacetone synthase is localized in the peroxisomal matrix of methanol-grownHansenula polymorpha.Arch. Microbiol. 143, 237–243 (1985).CrossRefGoogle Scholar
  3. Digan M.E., Lair S.V., Brierley R.A., Siegel R.S., Williams M.E., Ellis S.B., Kellaris P.A., Provow W.A., Craig W.S., Velicelebi G., Harpold M.M., Thill G.P.: Continuous production of a novel lysozyme via secretion from the yeast.Pichia pastoris. Bio/Technol. 7, 160–164 (1989).CrossRefGoogle Scholar
  4. Lam F.L., Lowande A.: The simultaneous assay of riboflavin 5-phosphate sodium salt and their water-soluble vitamins in liquid multivitamin formulations by liquid chromatography.J. Pharmac. Biomed. Anal. 6, 87–95 (1988).CrossRefGoogle Scholar
  5. Lück H.: Catalase, pp. 885–894 inMethods of Enzymatic Analysis (H.V. Bergmeyer, Ed.). Academic Press, New York-London 1963.Google Scholar
  6. Nicolay K., Veenhuis M., Douma A.C.: A 31P NMR study of the internal pH of yeast peroxisomes.Arch. Microbiol. 147, 37–41 (1987).PubMedCrossRefGoogle Scholar
  7. Reynolds E.S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.J. Cell. Biol. 17, 208–212 (1963).PubMedCrossRefGoogle Scholar
  8. Roggenkamp R., Sahm H., Hinkelmann W., Wagner F.: Alcohol oxidase and catalase in peroxisomes of methanol-grownCandida boidinii.Eur. J. Biochem. 59, 231–236 (1975).PubMedCrossRefGoogle Scholar
  9. Smith G., Tannhauser S.J.: A method for the determination of deoxyribonucleic acid, ribonucleic acid and phosphoproteins in animal tissue.J. Biol. Chem. 161, 83 (1945).Google Scholar
  10. Sreekrishna K., Potenz R.H.B., Cruze J.A., McCombie W.R., Parker K.A., Nelles L., Mazzaferro P.K., Holden K.A., Harrison R.G., Wood P.J., Phelps D.A., Hubbard C.E., Fuke M.: High level expression of heterologous proteins in methylotrophic yeastPichia pastoris.J. Basic. Microbiol. 28, 265–278 (1988).PubMedCrossRefGoogle Scholar
  11. Tani Y., Miya T., Nishikawa H., Ogata K.: The microbial metabolism of methanol. Part 1. Formation and crystalization of methanol-oxidizing enzyme in a methanol utilizing yeast,Kloeckera sp. no.2201.Agr. Biol. Chem. 36, 68–75 (1972).Google Scholar
  12. Veenhuis M., van Dijken J.P., Pilon S.A.F., Harder W.: Development of crystalline peroxisomes in methanol-grown cells of the yeastHansenula polymorpha and its relation to environmental conditions.Arch. Microbiol. 117, 153–163 (1978).PubMedCrossRefGoogle Scholar
  13. Veenhuis M., Harder W., van Dijken J.P., Mayer F.: Substructure of crystalline peroxisomes in methanol-grownHansenula polymorpha: evidence for anin vivo crystal of alcohol oxidase.Mol. Cell. Biol. 1, 949–954 (1981).PubMedGoogle Scholar
  14. Volfová O.: Studies on methanol-oxidizing yeasts. III. Enzymes.Folia Microbiol. 20, 307–319 (1975).CrossRefGoogle Scholar
  15. Volfová O., Kmentová M., Kyslíková E.: Acidotolerant strainCandida boidinii 2 utilizing methanol—characterization and biomass composition. (In Czech)Kvas. prům. 34, 330–331 (1988).Google Scholar
  16. Volfová O., Pilát P.: Studies on methanol-oxidizing yeast. I. Isolation and growth studies.Folia Microbiol. 19, 249–256 (1974).Google Scholar

Copyright information

© Folia Microbiologica 1992

Authors and Affiliations

  • O. Volfová
    • 1
  • M. Anděrová
    • 1
  • Z. Žižka
    • 1
  1. 1.Institute of MicrobiologyCzechoslovak Academy of SciencesPrague 4

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