Skip to content
1887

Microbiology Society logo Microbiology

Volume 129, Issue 5

Regulation of Enzyme Synthesis and Variation of Residual Methanol Concentration during Carbon-limited Growth of Kloeckera sp. 2201 on Mixtures of Methanol and Glucose Free

Abstract

The methylotrophic yeast sp. 2201 was grown in double carbon (glucose/methanol) limitation in a chemostat at a constant dilution rate and the regulation of the synthesis of enzymes involved in the assimilation of methanol and the breakdown of glucose was studied as a function of the composition of the glucose/methanol mixture in the inflowing medium. Enhanced synthesis of most of the enzymes taking part in the assimilation of C units took place despite the presence of glucose in the medium. Depending on the enzyme, 40 to 60% (w/w of total substrate) methanol in the mixture was enough to cause maximal induction and no further enhancement of their specific activities was observed during growth on mixtures containing higher methanol/glucose ratios. The only two enzymes involved in the assimilation of methanol not showing these characteristic patterns were classical transketolase and transaldolase. substrate fluxes through the individual steps of glucose and methanol metabolism have been calculated and compared with the experimentally determined specific activities of the enzymes concerned. The three enzymes whose specific activities were least in excess of the substrate flux calculated to be catalysed by them were transketolase, transaldolase and (possibly) dihydroxyacetone synthase.

The residual concentration of methanol was measured in the culture as a function of the composition of the supplied glucose/methanol mixture. During growth with mixtures the concentration of methanol was always lower than during growth with this substrate as the only carbon source. Possible ecological advantages accruing to a facultative methylotroph as a consequence of this behaviour are discussed.

  • Received:
  • Published Online:
© Society for General Microbiology, 1983
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-5-1269
1983-05-01
2025-05-01
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/5/mic-129-5-1269.html?itemId=/content/journal/micro/10.1099/00221287-129-5-1269&mimeType=html&fmt=ahah

References

  1. Amachi T., Bowien B. 1979; Characterisation of two fructose bisphosphataseisoenzymes from the hydrogen bacterium Nocardiaopuca lb. Journal of General Microbiology 113:347–356
     [Google Scholar]
  2. Attwood M. M., Van Dijken J. P. 1982; Characteristics of fructose-1,6-bisphosphatase from the methanol-utilizing yeast Hansenuia polymorpha. Journal of General Microbiology 128:2313–2317
     [Google Scholar]
  3. Babel W., Loffhagen N. 1979; Assimilation of methanol by yeasts, a new approach. Zeitschrift für allgemeine Mikrobiologie 19:299–302
     [Google Scholar]
  4. Bailey Y. E., Ollis D. F. 1977 Biochemical Engineering Fundamentals p. 349 New York: McGraw-Hill;
     [Google Scholar]
  5. Bücher T., Pfleiderer G. 1955; Pyruvate kinase from muscle. Methods in Enzymotogy 1:435–440
     [Google Scholar]
  6. Bull A. T., Brown C. M. 1979; Continuous culture applications to microbial biochemistry. In Microbial Biochemistry. International Review of Biochemistry 21 pp. 177–226 Edited by Quayle J. R. Baltimore: University Park Press;
     [Google Scholar]
  7. Van Dijken J. P., Quayle J. R. 1977; Fructose metabolism in four Pseudomonas species. Archives of Microbiology 114:281–286
     [Google Scholar]
  8. Van Dijken J.P, Harder , Beardsmore A. J., Quayle J. R. 1978; Dihydroxyacetone: an intermediate in the assimilation of methanol by yeasts?. EEMS Microbiology Letters 4:97–102
     [Google Scholar]
  9. Eggeling L., Sahm H. 1980; Regulation of alcohol oxidase synthesis in Hansenuia polymorpha:oversynthesis during growth on mixed substrates and induction by methanol. Archives of Microbiology 127:119–124
     [Google Scholar]
  10. Eggeling L., Sahm H. 1981; Enhanced utilization of methanol during growth on a mixed substrate: a continuous culture study with Hansenuia polymorpha. Archives of Microbiology 130:362–365
     [Google Scholar]
  11. Egli T. 1980 Wachstum von Methanol-assimilieren-den Hefen. Eine Chemostat-Studie über die Regulation der Methanol-dissimilierenden Enzyme. Dissertation no. 6538, ETH, Zürich, Switzerland
    [Google Scholar]
  12. Egli T., Fiechter A. 1981; Theoretical analysis of media used in the growth of yeasts on methanol. Journal of General Microbiology 123:365–369
     [Google Scholar]
  13. Egli T., Käppeli O. 1980; Simultaneous utilisation of glucose and methanol in chemostat cultures of Hansenulapolymorpha and Kloeckera sp. 2201. 3rd International Symposium on Microbial Growth on C, - Compounds, Sheffield, U.K. (Abstract p. 19)
    [Google Scholar]
  14. Egli T., Van Dijken J. P., Veenhuis M., Harder W., Fiechter A. 1980; Methanol metabolism in yeasts: regulation of the synthesis of catabolic enzymes. Archives of Microbiology 124:115–121
     [Google Scholar]
  15. Egli T., Käppeli O., Fiechter A. 1982a; Regulatory flexibility of methylotrophic yeasts in chemostat cultures: simultaneous assimilation of glucose and methanol at a fixed dilution rate. Archives of Microbiology 131:1–7
     [Google Scholar]
  16. Egli T., Käppeli O., Fiechter A. 1982b; Mixed substrate growth of methylotrophic yeasts in chemostat culture: influence of the dilution rate on the utilisation of a mixture of glucose and methanol. Archives of Microbiology 131:8–13
     [Google Scholar]
  17. Gottschal J. C., De Vries S., Kuenen J. G. 1979; Competition between the facultative chemo- lithotrophicThiobacillus A2, an obligately chemolithotrophic Thiobacillus and a heterotrophic Spirillum for inorganic and organic substrates. Archives of Microbiology 121:241–248
     [Google Scholar]
  18. Harder W., Dijkhuizen L. 1976; Mixed substrate utilization in microorganisms. In Continuous Culture 6: Applications and New Fields pp. 297–314 Edited by Dean A. C. R., Ellwood D. C., Evans C. G. T., Melling T. Chichester & Oxford: Ellis Horwood;
     [Google Scholar]
  19. Harder W., Dijkhuizen L. 1982; Strategies of mixed substrate utilization in microorganisms. In New Dimensions in Microbiology: Mixed substrates, Mixed Cultures and Microbial Communities pp. 459–480 Edited by Quayle J. R., Bull A. T. London: Royal Society;
     [Google Scholar]
  20. Harder W., Van Dijken J. P. 1975; A theoretical study of growth yields of yeasts on methanol. In Microbial Growth on C1-Compounds pp. 155–161 Tokyo: Society of Fermentation Technology;
     [Google Scholar]
  21. Kato N., Nishizawa T., Sakazawa C., Tani Y., Yamada H. 1979; Xylulose 5-phosphate-dependent fixation of formaldehyde in a methanolutilizing yeast, Kloeckera sp. 2201. Agricultural and Biological Chemistry 43:2013–2015
     [Google Scholar]
  22. Law A. T., Button D. K. 1977; Multiple-carbon source limited growth kinetics of a marine coryne- form bacterium. Journal of Bacteriology 129:115–123
     [Google Scholar]
  23. Law A. T., Robertson B. R., Dunker S. S., Button D. K. 1976; On describing microbial growth kinetics from continuous culture data: some general considerations, observations and concepts. Microbial Ecology 2:261–283
     [Google Scholar]
  24. Lee J.-D., Komagata K. 1980; Taxonomic study of methanol-assimilating yeasts. Journal of General and Applied Microbiology 26:133–158
     [Google Scholar]
  25. Lindley N. D., Waites M. J., Quayle J. R. 1980; A modified pulse-labelling technique for the detection of early intermediates in microbial metabolism: detection of [l4C]dihydroxyacetone during assimilation of [l4C]methanol by Hansenula polymorpha. FEMS Microbiology Letters 8:13–16
     [Google Scholar]
  26. Von Meyenburg K. 1969; Katabolit-Repression und der Sprossungszyklus von Saccharomyces cerevisiae. Dissertation no. 4279, ETH, Zurich, Switzerland
    [Google Scholar]
  27. Ng F.M.-W., Dawes E. A. 1973; Chemostat studies on the regulation of glucose metabolism in Pseudomonas aeruginosa by citrate. Biochemical Journal 132:129–140
     [Google Scholar]
  28. O’Connor M., Quayle J. R. 1979; Mutants of Hansenula polymorpha and Candida boidinii impaired in their ability to grow on methanol. Journal of General Microbiology 113:203–208
     [Google Scholar]
  29. Powell E. O. 1967; The growth rate of microorganisms as a function of substrate concentration. In Microbial Physiology and Continuous Culture. Proceedings of the 3rd International Symposium on Continuous Culture of Microorganisms pp. 34–55 Edited by Powell E. O., Evans C. G. T., Strange R. E., Tempest D. W. London: H.M.S.O;
     [Google Scholar]
  30. Ratledge C., Botham P. A. 1977; Pathways of glucose metabolism in Candida 107, a lipid-accumulating yeast. Journal of General Microbiology 102:391–395
     [Google Scholar]
  31. Rittenberg S. C. 1972; The obligate autotroph - the demise of a concept. Antonie van Leeuwenhoek 38:457–478
     [Google Scholar]
  32. Schlegel H. G. 1981 Allgemeine Mikrobiologie p. 223 Stuttgart: Georg Thieme Verlag;
     [Google Scholar]
  33. Tchola O., Horecker B. L. 1966; Transaldolase. Methods in Enzymology 9:499–505
     [Google Scholar]
  34. Waites M. J., Quayle J. R. 1980; Dihydroxyacetone: a product of xylulose 5-phosphate-dependent fixation of formaldehyde by methanol-grown Candida boidinii. Journal of General Microbiology 118:321–327
     [Google Scholar]
  35. Waites M. J., Quayle J. R. 1981; The interrelation between transketolase and dihydroxyacetone synthase activities in the methylotrophic yeast Candida boidinii. Journal of General Microbiology 124:309–316
     [Google Scholar]
  36. Waites M. J., Quayle J. R. 1983; Dihydroxyacetone synthase: a special transketolase for formaldehyde fixation from the methylotrophic yeast Candida boidinii CBS 5777. Journal of General Microbiology 129:935–944
     [Google Scholar]
  37. Waites M. J., Lindley N. D., Quayle J. R. 1981; Determination of the labelling pattern of dihydroxyacetone and hexose phosphate following a brief incubation of methanol-grown Hansenulapolymorpha with [14C]methanol. Journal of General Microbiology 122:193–199
     [Google Scholar]
  38. Wood T. 1970; Spectrophotometric assay for d-ribose-5-phosphate ketol-isomerase and for d-ribulose-5-phosphate 3-epimcrase. Analytical Biochemistry 33:297–306
     [Google Scholar]
/content/journal/micro/10.1099/00221287-129-5-1269
Loading
Regulation of Enzyme Synthesis and Variation of Residual Methanol Concentration during Carbon-limited Growth of Kloeckera sp. 2201 on Mixtures of Methanol and Glucose
Microbiology 129, 1269 (1983); https://doi.org/10.1099/00221287-129-5-1269
/content/journal/micro/10.1099/00221287-129-5-1269
/content/journal/micro/10.1099/00221287-129-5-1269
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error