Archives of Microbiology

, Volume 150, Issue 2, pp 155–159 | Cite as

Dihydroxyacetone kinase from a methylotrophic yeast, Hansenula polymorpha CBS 4732

Purification, characterization and physiological role
  • Nobuo Kato
  • Hiroshi Yoshikawa
  • Katsuhiko Tanaka
  • Masayuki Shimao
  • Chikahiro Sakazawa
Original Papers


Dihydroxyacetone (DHA) kinase was purified to electrophoretic homogeneity from methanol-grown Hansenula polymorpha CBS 4732. The enzyme was a dimer with a molecular weight of 150,000, and had an isoelectric point of 4.9. The enzyme was active toward DHA, and D- and L-glyceraldehydes as phosphorylation acceptors, and only ATP served as a donor. ADP inhibited the enzyme at a physiological concentration. Magnesium ion was essential for the activity and stability. Some other divalent cations can substitute in part the magnesium ion. The DHA kinases found in cells grown on methanol and glycerol were immunologically identical, but were different from those of other methylotrophic yeasts as shown by immunotitration. A mutant (204D) derived from the yeast, which could not grow on methanol or DHA but could so on glycerol, was deficient in DHA kinase. Glycerol kinase activity was found in glycerol-grown 204D cells as well as the parent strain.

Key words

Hansenula polymorpha Dihydroxyacetone kinase Glycerol kinase Methanol Glycerol 





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  1. Babel W, Hofmann KH (1982) The relation between the assimilation of methanol and glycerol in yeasts. Arch Microbiol 127:119–124Google Scholar
  2. Bystrykh LV, Trotsenko YA (1983) Purification and properties of dihydroxyacetone kinase from methylotrophic yeast, Candida boidinii. Biokhimiya 48:1611–1616Google Scholar
  3. Dijken JP van, Harder W, Beardsmore AJ, Quayle JR (1978) Dihydroxyacetone: an intermediate in the assimilation of methanol by yeast? FEMS Microbiol Lett 4:97–102Google Scholar
  4. Dixon M, Webb EC (1979) Enzymes, 3rd ed. Longman Group Ltd, LondonGoogle Scholar
  5. Garvey JS, Cremer NE, Sussforf DH (1977) Methods in Immunology, a laboratory text for instruction and research, 3rd ed. W. A. Benjamin Inc, MassachusettsGoogle Scholar
  6. Hofmann KH, Babel W (1982) Dihydroxyacetone kinase of methanol-assimilating yeasts. II. Partial purification and some properties of dihydroxyacetone kinase from Candida methylica. Z Allg Mikrobiol 21:219–224Google Scholar
  7. Johnson EA, Burke SK, Forage RG, Lin ECC (1984) Purification and properties of dihydroxyacetone kinase from Klebsiella pneumonia. J Bacteriol 160:55–60Google Scholar
  8. 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. 2201. Biochim Biophys Acta 715:143–150Google Scholar
  9. Kato N, Kobayashi H, Shimao M, Sakazawa C (1984) Properties of formaldehyde dismutation catalyzing enzyme of Pseudomonas putida F61. Agric Biol Chem 48:2017–2023Google Scholar
  10. Kato N, Kobayashi H, Shimao M, Sakazawa C (1986a) Dihydroxyacetone production from methanol by a dihydroxyacetone kinase deficient mutant of Hansenula polymorpha. Appl Microbiol Biotechnol 23:180–186Google Scholar
  11. Kato N, Yamagami T, Shimao M, Sakazawa C (1986b) Formaldehyde dismutase, a novel NAD-binding oxidoreductase from Pseudomonas putida F61. Eur J Biochem 156:59–64Google Scholar
  12. Koning W de, Gleeson MAG, Harder W, Dijkhuizen L (1987a) Regulation of methanol metabolism in the yeast Hansenula polymorpha. Arch Microbiol 147:375–382Google Scholar
  13. Koning W de, Harder W, Dijkhuizen L (1987b) Glycerol metabolism in the methylotrophic yeast Hansenula polymorpha: phosphorylation as the initial step. Arch Microbiol 148:314–320Google Scholar
  14. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227:680–685Google Scholar
  15. Lerner HR, Sussman I, Avron M (1980) Characterization and partial purification of dihydroxyacetone kinase in Dunaliella salina. Biochim Biophys Acta 615:1–9Google Scholar
  16. Levine DW, Cooney CL (1973) Isolation and characterization of a thermotolerant methanol-utilizing yeast. Appl Microbiol 26:982–990Google Scholar
  17. Lin ECC (1976) Glycerol dissimilation and its regulation in bacteria. Annu Rev Microbiol 30:535–578Google Scholar
  18. Livingston DM (1974) Immunoaffinity chromatography of proteins. In: Jakoby WB, Wilchek M (eds) Methods in enzymology, vol 34. Academic Press, New York San Francisco London, pp 723–731Google Scholar
  19. Marshal JH, May JW, Sloan J, Vasiliadis GE (1986) Purification and properties of dihydroxyacetone kinase from Schizosaccharomyces pombe. J Gen Microbiol 132:2611–2614Google Scholar
  20. O'Conner ML, Quayle JR (1979) Mutants of Hansenula polymorpha and Candida boidinii and in their ability to grow on methanol. J Gen Microbiol 113:203–208Google Scholar
  21. Polakis ES, Bartley W (1966) Changes in the intracellular concentrations of adenosine phosphates and nicotinamide nucleotides during the aerobic growth cycle of yeast on different carbon sources. Biochem J 99:521–533Google Scholar
  22. Tachiki T, Chigiri E, Tochikura T (1987) Purification and properties of dihydroxyacetone kinase from Gluconobacter suboxydans. J Ferment Technol 65:107–110Google Scholar
  23. Tani Y, Yamada K (1987a) Diversity in glycerol metabolism of methylotrophic yeasts. FEMS Microbiol Lett 40:151–153Google Scholar
  24. Tani Y, Yamada K (1987b) Glycerol metabolism of methylotrophic yeasts. Agric Biol Chem 51:1927–1933Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Nobuo Kato
    • 1
  • Hiroshi Yoshikawa
    • 1
  • Katsuhiko Tanaka
    • 1
  • Masayuki Shimao
    • 1
  • Chikahiro Sakazawa
    • 1
  1. 1.Department of Environmental Chemistry and TechnologyTottori UniversityTottoriJapan

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