1887

Abstract

Nitrogen assimilation during growth of on methylated amines as sole nitrogen source involves NADP-dependent glutamate dehydrogenase. Changes in enzyme activities during the adaptation of the yeast from growth on ammonium to growth on trimethylamine were examined. No ammonia, dimethylamine or monomethylamine could be detected in the medium during growth on trimethylamine. When two methylated amines were supplied together, they were used simultaneously, although monomethylamine was metabolized more quickly than the others. When cells were grown on a low concentration of ammonium plus higher concentrations of di- or trimethylamine, the ammonium was used first. NADP-dependent glutamate dehydrogenase was the first enzyme to be derepressed, followed by methylamine oxidase and formaldehyde dehydrogenase. Di- and trimethylamine mono-oxygenase activities only appeared when the ammonium concentration fell below 0.5 mM. At this point amine utilization could be detected and no diauxic lag was observed in the growth curve. During growth on limiting ammonium, there was an increase in the activity of methylamine oxidase (150-fold) and catalase (5-fold) in the absence of any amine, but no amine mono-oxygenase activity was detected. Addition of ammonium ions to cultures growing on dimethylamine produced an immediate repression of synthesis of methylamine oxidase, NADP-dependent glutamate dehydrogenase and the two amine mono-oxygenases. An inverse correlation was found between intracellular ammonium concentration and methylamine oxidase activity. Ammonium ions also inhibited the uptake of dimethylamine or trimethylamine by washed suspensions of dimethylamine-grown cells. It is concluded that the control of methylamine oxidase and catalase and (independently) of NADP-dependent glutamate dehydrogenase is by repression of enzyme synthesis by ammonium, while expression of amine mono-oxygenases seems to require the amine to be present in the medium. Formaldehyde and formate dehydrogenases seem also to be induced by their respective substrates.

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1984-08-01
2021-10-21
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References

  1. Bradford M. M. 1976; A rapid and sensitive method for the quantitative determination of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  2. Brown C. M. 1976; Nitrogen metabolism in bacteria and fungi. In Continuous Culture 6 Applications and New Fields pp. 170–183 Edited by Dean A. C. R., Ellwood D. C., Evans C. G. T., Melling J. Chichester: Ellis Horwood;
    [Google Scholar]
  3. Bruinenberg P. M., Van Dijken J. P., Scheffers WA. 1983; An enzymic analysis of NADPH production and consumption in Candida utilis. Journal of General Microbiology 129:965–971
    [Google Scholar]
  4. Chaney AL., Marbach E. P. 1962; Modified reagents for determination of urea and ammonia. Clinical Chemistry 8:130–132
    [Google Scholar]
  5. Dalton H. 1979; Utilization of inorganic nitrogen by microbial cells. In Microbial Biochemistry International Review of Biochemistry 21 pp. 227–266 Edited by Quayle J. R. Baltimore: University Park Press;
    [Google Scholar]
  6. Van Dijken J. P., Bos P. 1981; Utilization of amines by yeasts. Archives of Microbiology 128:320–324
    [Google Scholar]
  7. Van Dijken J. P., Harder W., Beardsmore AJ., Quayle J. R. 1978; Dihydroxyacetone: an intermediate in the assimilation of methanol by yeasts?. FEMS Microbiology Letters 4:97–102
    [Google Scholar]
  8. Doherty D. 1970; l-Glutamate dehydrogenases (yeast). Methods in Enzymology 17A:850–856
    [Google Scholar]
  9. Dubin D. T. 1960; The assay and characterization of amines by means of 2,4-dinitrofluorobenzene. Journal of Biological Chemistry 235:783–786
    [Google Scholar]
  10. Green J., Large P. J. 1983a; Oxidation of dimethylamine and trimethylamine in methazotro- phic yeasts by microsomal mono-oxygenases sensitive to carbon monoxide. Biochemical and Biophysical Research Communications 113:900–907
    [Google Scholar]
  11. Green J., Large P. J. 1983b; Cell-free oxidation of dimethylamine by a mono-oxygenase in the methazotrophic yeast Candida utilis. Biochemical Society Transactions 11:786
    [Google Scholar]
  12. Green J, Large P. J. 1982; More than one amine oxidase is involved in the metabolism by yeasts of primary amines supplied as nitrogen source. Journal of General Microbiology 128:991–996
    [Google Scholar]
  13. Green J., Haywood G. W., Large P. J. 1983; Serological differences between the multiple amine oxidases of yeasts and comparison of the specificities of the purified enzymes from Candida utilis and Pichia pastoris. Biochemical Journal 211:481–493
    [Google Scholar]
  14. Haywood G. W., Large P. J. 1981; Microbial oxidation of amines. Distribution, purification and properties of two primary-amine oxidases from the yeast Candida boidinii grown on amines as sole nitrogen source. Biochemical Journal 199:187–201
    [Google Scholar]
  15. Haywood G. W., Large P. J. 1984; Partial purification of a peroxisomal polyamine oxidase from Candida boidinii and its role in growth on spermidine as sole nitrogen source. Journal of General Microbiology 130:1123–1136
    [Google Scholar]
  16. Lang E., Lang H. 1972; Spezifische Farbreaktion zum direkten Nachweis der Ameisensaure. Zeitschrift für analytische Chemie 260:8–10
    [Google Scholar]
  17. Large P. J., Green J. 1984; Oxidation of mono-, di- and trimethylamine by methazotrophic yeasts: properties of the microsomal and peroxisomal enzymes involved and comparison with bacterial systems. In Proceedings of Fourth International Symposium on Microbial Growth on C1 Compounds Edited by Crawford R. L. Washington: American Society for Microbiology (in the Press);
    [Google Scholar]
  18. Large P. J., Mcdougall H. 1975; An enzymic method for the micro-estimation of trimethylamine. Analytical Biochemistry 64:304–310
    [Google Scholar]
  19. Large P. J., Eady R. R., Murden D. J. 1969; An enzymic method for the micro-estimation of methyl- amine, ethylamine, and n-propylamine. Analytical Biochemistry 32:402–407
    [Google Scholar]
  20. Meiberg J. B. M, Harder W. 1979; Dimethylamine dehydrogenase from Hyphomicrobium X. Purification and some properties of a new enzyme that oxidizes secondary amines. Journal of General Microbiology 115:49–58
    [Google Scholar]
  21. Nash T. 1953; The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochemical Journal 55:416–421
    [Google Scholar]
  22. O’Connor M. L., Quayle J. R. 1980; Pentose phosphate-dependent fixation of formaldehyde by methanol-grown Hansenula polymorpha and Candida boidinii. Journal of General Microbiology 120:219–225
    [Google Scholar]
  23. Roon R. J., Murdoch M., Kunze B., Dunlop P. C. 1982; Derepression of asparaginase II during exponential growth of Saccharomyces cerevisiae on ammonium ion. Archives of Biochemistry and Biophysics 219:101–109
    [Google Scholar]
  24. Rowe W. B., Ronzio R. A., Wellner V. P., Meister A. 1970; Glutamine synthetase (sheep brain). Methods in Enzymology 17A:900–910
    [Google Scholar]
  25. Schütte H., Flossdorf J., Sahm H., Kula M.-R. 1976; Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii. European Journal of Biochemistry 62:151–160
    [Google Scholar]
  26. Veenhuis M., Zwart K. B.jr 1981; Biogenesis and turnover of peroxisomes involved in the concurrent oxidation of methanol and methyl- amine in Hansenula polymorpha. Archives of Microbiology 129:35–41
    [Google Scholar]
  27. Werner W., Rey H.-G., Wielinger H. 1970; Über die Eigenschaften eines neuen Chromogens für die Blutzuckerbestimmung nach der GOD/POD- Methode. Zeitschrift für analytische Chemie 252:224–228
    [Google Scholar]
  28. Zwart K. B. 1983 Metabolic significance of microbodies in the yeasts Candida utilis and Hansenula polymorpha Doctoral thesis University of Groningen; The Netherlands:
    [Google Scholar]
  29. Zwart K. B., Harder W. 1983; Regulation of the metabolism of some alkylated amines in the yeasts Candida utilis and Hansenula polymorpha. Journal of General Microbiology 129:3157–3169
    [Google Scholar]
  30. Zwart K., Veenhuis M., Van Dijken J. P., Harder W. 1980; Development of amine-oxidase- containing peroxisomes in yeasts during growth on glucose in the presence of methylamine as the sole source of nitrogen. Archives of Microbiology 126:117–126
    [Google Scholar]
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