Skip to main content
Log in

Regulation of methanol metabolism in the yeast Hansenula polymorpha

Isolation and characterization of mutants blocked in methanol assimilatory enzymes

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

A study of enzyme profiles in Hansenula polymorpha grown on various carbon substrates revealed that the synthesis of the methanol dissimilatory and assimilatory enzymes is regulated in the same way, namely by catabolite repression and induction by methanol. Mutants of H. polymorpha blocked in dihydroxyacetone (DHA) synthase (strain 70 M) or DHA kinase (strain 17 B) were unable to grow on methanol which confirmed the important role attributed to these enzymes in the biosynthetic xylulose monophosphate (XuMP) cycle. Both mutant strains were still able to metabolize methanol. In the DNA kinase-negative strain 17 B this resulted in accumulation of DHA. Although DHA kinase is thought to be involved in DHA and glycerol metabolism in methylotrophic yeasts, strain 17 B was still able to grow on glycerol at a rate similar to that of the wild type. DHA on the other hand only supported slow growth of this mutant when relatively high concentrations of this compound were provided in the medium. This slow but definite growth of strain 17 B on DHA was not based on the reversible DHA synthase reaction but on conversion of DHA into glycerol, a reaction catalyzed by DNA reductase. The subsequent metabolism of glycerol in strain 17 B and in wild type H. polymorpha, however, remains to be elucidated.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

XuMP:

xylulose monophosphate

DHA:

dihydroxyacetone

EMS:

ethyl methanesulphonate

References

  • Avigad G (1983) A simple spectrophotometric determination of formaldehyde and other aldehydes: Application to periodateoxidized glycolsystems. Anal Biochem 134:499–504

    PubMed  Google Scholar 

  • Babel W, Hofmann KH (1982) The relation between the assimilation of methanol and glycerol in yeasts. Arch Microbiol 132:179–184

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Brooke AG, Dijkhuizen L, Harder W (1986) Regulation of flavin biosynthesis in the methylotrophic yeast Hansenula polymorpha. Arch Microbiol 145:62–70

    Google Scholar 

  • Bublitz C, Wieland O (1962) Glycerokinase. In: Colowick SP, Kaplan NO (eds) Methods enzymology, vol 5. Academic Press, New York, pp 354–361

    Google Scholar 

  • Bystrykh LV, Sokolov AP, Trotsenko YA (1981) Purification and properties of dihydroxyacetone synthase from the methylotrophic yeast Candida boidinii. FEBS Lett 132:324–328

    Google Scholar 

  • Bystrykh LV, Trotsenko YA (1983) Purification and some properties of dihydroxyacetone kinase from the methylotrophic yeast Candida boidinii. Biochemistry (USSR) 48:1611–1616

    Google Scholar 

  • Bystrykh LV (1985) Kinetic properties of dihydroxyacetone kinase of the methylotrophic yeast Candida boidinii KD1. Biochemistry (USSR) 50:1012–1017

    Google Scholar 

  • Dijken JP van, Harder W, Beardsmore AJ, Quayle JR (1978) Dihydroxyacetone: an intermediate in the assimilation of methanol by yeasts? FEMS Microbiol Lett 4:97–102

    Google Scholar 

  • Dijken JP van, Otto R, Harder W (1976a) Growth of Hansenula polymorpha in a methanol-limited chemostat. Physiological responses due to the involvement of methanol oxidase as a key enzyme in methanol metabolism Arch Microbiol 111:137–144

    PubMed  Google Scholar 

  • Dijken JP van, Oostra-Demkes GJ, Otto R, Harder W (1976b) S-Formylglutathione: the substrate for formate dehydrogenase in methanol-utilizing yeasts. Arch Microbiol 111:77–83

    PubMed  Google Scholar 

  • Eggeling L, Sahm H (1978) Derepression and partial insensitivity to carbon catabolite repression of the methanol dissimilating enzymes in Hansenula polymorpha. Eur J Appl Microbiol Biotechnol 5:197–202

    Google Scholar 

  • Egli T, Dijken JP van, Veenhuis M, Harder W, Fiechter A (1980) Methanol metabolism in yeasts: regulation of the synthesis of catabolic enzymes. Arch Microbiol 124:115–121

    Google Scholar 

  • Egli T, Lindley ND, Quayle JR (1983) Regulation of enzyme synthesis and variation of residual methanol concentration during carbon-limited growth of Kloeckera sp. 2201 on mixtures of methanol and glucose. J Gen Microbiol 129:1269–1281

    Google Scholar 

  • Gleeson MA, Waites MJ, Sudbery PE (1984) Development of techniques for genetic analysis in the methylotrophic yeast Hansenula polymorpha. In: Crawford RL, Hanson RS (eds) Microbial growth on C1 compounds. American Society for Microbiology, Washington DC, pp 228–235

    Google Scholar 

  • Harder W, Trotsenko YA, Bystrykh LV, Egli T (1987) Metabolic regulation in methylotrophic yeasts. In: van Verseveld HW, Duine JA (eds) Microbial growth on C1 compounds. Martinus Nijhoff, Dorderecht, pp 139–149

    Google Scholar 

  • Hofmann KH, Babel W (1980) Dihydroxyacetone kinase of methanol-assimilating yeasts. I. Regulation of dihydroxyacetone kinase from Candida methylica in situ. Z Allg Mikrobiol 20:389–398

    PubMed  Google Scholar 

  • Hofmann KH, Babel W (1981) Dihydroxyacetone kinase of methanol-assimilating yeasts. II. Partial purification and some properties of dihydroxyacetone kinase from Candida methylica. Z Allg Mikrobiol 21:219–224

    PubMed  Google Scholar 

  • Hofmann K (1983) Assimilation und Akkumulation von Glycerol durch Hefen und Schimmelpilze. Biol Rdsch 21:265–276

    Google Scholar 

  • Kato N, Higuchi T, Sakazawa C, Nishizawa T, Tani Y, Yamada H (1982) Purification and properties of a transketolase responsible for formaldehyde fixation in a methanol-utilizing yeast Candida boidinii (Kloeckera sp.) No. 2011. Biochim Biophys Acta 715:143–150

    PubMed  Google Scholar 

  • Kato N, Kobayashi H, Shimao M, Sakazawa C (1986) Dihydroxyacetone production from methanol by a dihydroxyacetone kinase deficient mutant of Hansenula polymorpha. Appl Microbiol Biotechnol 23:180–186

    Google Scholar 

  • Koning W de, Dijkhuizen L (1986) The relationship between methanol, dihydroxyacetone and glycerol metabolism in the methylotrophic yeast Hansenula polymorpha. Abstr. 5th Int Symp on Microbial Growth on C1 Compounds. Haren, The Netherlands, p 91

  • Laanbroek HJ, Abee T, Woogd JL (1982) Alcohol conversions by Desulfobulbus propionicus Lindhorst in the presence and absence of sulfate and hydrogen. Arch Microbiol 133:178–184

    Google Scholar 

  • O'Connor ML, Quayle JR (1979) Mutants of Hansenula polymorpha and Candida boidinii impaired in their ability to grow on methanol. J Gen Microbiol 113:203–208

    Google Scholar 

  • Trotsenko YA, Bystrykh LV (1985) Regulation of primary pathways of methanol metabolism in methylotrophic yeasts. In: Kulaev IS, Dawes EA, Tempest DW (eds) Environmental regulation of microbial metabolism. Academic Press, London, pp 113–120

    Google Scholar 

  • Veenhuis M, Douma A, Harder W, Osumi M (1983) Degradation and turnover of peroxisomes in the yeast Hansenula polymorpha induced by selective inactivation of peroxisomal enzymes. Arch Microbiol 134:193–203

    PubMed  Google Scholar 

  • Vishniac W, Santer M (1957) The thiobacilli. Bacteriol Rev 21:195–213

    PubMed  Google Scholar 

  • Waites MJ, Quayle JR (1981) The interrelation between transketolase and dihydroxyacetone synthase activities in the methylotrophic yeast Candida boidinii. J Gen Microbiol 124:309–316

    PubMed  Google Scholar 

  • Waites MJ, Quayle JR (1983) Dihydroxyacetone synthase: a special transketolase for formaldehyde fixation from the methylotrophic yeast Candida boidinii CBS 5777. J Gen Microbiol 129:935–944

    Google Scholar 

  • Wieland O (1974) Glycerol. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 3. Academic Press, New York, pp 1404–1409

    Google Scholar 

  • Zwart KB, Harder W (1983) Regulation of the metabolism of some alkylated amines in the yeasts Candida utilis and Hansenula polymorpha. J Gen Microbiol 129:3157–3169

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

de Koning, W., Gleeson, M.A.G., Harder, W. et al. Regulation of methanol metabolism in the yeast Hansenula polymorpha . Arch. Microbiol. 147, 375–382 (1987). https://doi.org/10.1007/BF00406136

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00406136

Key words

Navigation