1887

Microbiology Society logo

Volume 114, Issue 2

A Biochemical Explanation for Lipid Accumulation in Candida 107 and Other Oleaginous Micro-organisms Free

Abstract

The biochemical explanation for lipid accumulation was investigated principally in 107 and, for comparison, in the non-oleaginous yeast There were no significant differences between these two yeasts in their control of glucose uptake; in both yeasts, the rates of glucose uptake were independent of the growth rate and were higher in carbon-limited chemostat cultures than in nitrogen-limited cultures. There was no lipid turnover in either yeast, as judged from [C]acetate uptake and subsequent loss of C from the lipid of steady-state chemostat cultures. Acetyl-CoA carboxylase from both yeasts was similar in most characteristics except that from 107 was activated by citrate (40% activation at 1 m). The enzyme from 107 was relatively unstable and, when isolated from nitrogen-limited (lipid-accumulating) cultures, was accompanied by a low molecular weight inhibitor.

The reason for lipid accumulation is attributed to the decrease in the intracellular concentration of AMP as cultures become depleted of nitrogen. As the NAD-dependentisocitrate dehydrogenase of 107, but not , requires AMP for activity, themetabolism of citrate through the tricarboxylic acid cycle in the mitochondria becomesarrested. In 107, but not in , there is an active ATP:citrate lyase whichconverts the accumulating citrate, when it passes into the cytosol, into acetyl-CoA and oxaloacetate. The former product is then available for fatty acid biosynthesis which is stimulated by the high ATP concentration within the cells, by the activation of acetyl-CoA carboxylase by citrate and by the provision of NADPH generated as oxaloacetate is converted via malate to pyruvate.

Similar characteristics were evident in oleaginous strains of and but not in non-oleaginous representatives of these species.

  • Received:
  • Published Online:
© Society for General Microbiology, 1979
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-114-2-361
1979-10-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/114/2/mic-114-2-361.html?itemId=/content/journal/micro/10.1099/00221287-114-2-361&mimeType=html&fmt=ahah

References

  1. Anfinsen C.B. 1955; Aconitase from pig heart muscle. Methods in Enzymology 1:695–698
     [Google Scholar]
  2. Atkinson D.E. 1969; Regulation of enzyme function. Annual Review of Microbiology 23:47–68
     [Google Scholar]
  3. Atkinson D.E. 1977 Cellular Energy Metabolism and its Regulation. New York, San Francisco & London: Academic Press;
     [Google Scholar]
  4. Babij T., Moss F.J., Ralph B.J. 1969; Effects of oxygen and glucose levels on lipid composition of the yeast Candida utilis growing in continuous culture. Biotechnology and Bioengineering 11:593–603
     [Google Scholar]
  5. Bernhauer K., Rauch J. 1948; Beitrage zur mikrobiologischen Eiweiss- und Fettsynthese. I. Die grundlegenden Bedingungen für die Eiweiss und Fett Produktion durch Mycelpilze in der Submers-Kultur. Biochemische Zeitschrift 319:77–93
     [Google Scholar]
  6. Botham P.A., Ratledge C. 1978; Metabolic studies related to lipid accumulation in yeast. Biochemical Society Transactions 6:383–385
     [Google Scholar]
  7. Briquet M. 1977; Transport of pyruvate and lactate in yeast mitochondria. Biochimica et biophysica acta 459:290–299
     [Google Scholar]
  8. Chapman A.G., Atkinson D.E. 1977; Adenine nucleotide concentrations and turnover rates.Their correlation with biological activity in bacteria and yeast. Advances in Microbial Physiology 15:253–306
     [Google Scholar]
  9. Chapman A.G., Fall L., Atkinson D.E. 1971; Adenylate energy charge in Escherichia coli during growth and starvation. Journal of Bacteriology 108:1072–1086
     [Google Scholar]
  10. Chapman C., Bartley W. 1969; Adenosine phosphates and the control of glycolysis and gluconeogenesis in yeast. Biochemical Journal 111:609–613
     [Google Scholar]
  11. Dawes E.A., Large P.J. 1970; Effect of starvation on the viability and cellular constituents of Zymomonas anaerobia and Zymomonas mobilis. Journal of General Microbiology 60:31–42
     [Google Scholar]
  12. Dolezal J., Kapralek F. 1976; Physiological characteristics of chemostatically grown Citrobacter freundii as a function of specific growth rate and type of nutrient limitation. Folia microbiologica 21:168–177
     [Google Scholar]
  13. Gill C. O., Ratledge C. 1973a; Inhibition of glucose assimilation and transport by n-decane and other n-alkanes in Candida 107. Journal of General Microbiology 75:11–22
     [Google Scholar]
  14. Gill C. O., Ratledge C. 1973b; Regulation of de novo fatty acid biosynthesis in the n-alkane utilizing yeast, Candida 107. Journal of General Microbiology 78:337–347
     [Google Scholar]
  15. Gill C.O., Hall M.J., Ratledge C. 1977; Lipid accumulation in an oleaginous yeast, Candida 107, growing on glucose in single-stage continuous culture. Applied and Environmental Microbiology 33:231–239
     [Google Scholar]
  16. Gornall A.G., Bardawill C.J., David M.M. 1949; Determination of serum proteins by means of the biuret reaction. Journal of Biological Chemistry 177:751–766
     [Google Scholar]
  17. Haarasilta S., Taskinen L. 1977; Location of three key enzymes of gluconeogenesis in baker’s yeast. Archives of Microbiology 113:159–161
     [Google Scholar]
  18. Hall M.J. 1978 Growth of oleaginous yeasts in continuous culture. M.Sc. Thesis, University of Hull.
    [Google Scholar]
  19. Harrison D.E.F., Maitra P.K. 1969; Control of respiration and metabolism in growing Klebsiella aerogenes. The role of adenine nucleotides. Biochemical Journal 112:123–134
     [Google Scholar]
  20. Hathaway J.A., Atkinson D.E. 1963; The effect of adenylic acid on yeast nicotinamide adenine dinucleotide isocitrate dehydrogenase, a possible metabolic control mechanism. Journal of Biological Chemistry 238:2875–2881
     [Google Scholar]
  21. Hayashi E., Hasegawa R., Tomita T. 1978; The fluctuation of various enzyme activities due to myoinositol deficiency in Saccharomyces cerevisiae. Biochimica et biophysica acta 540:231–237
     [Google Scholar]
  22. Hobson P.N., Summers R. 1972; ATP pool and growth yield in Selenomonas ruminantium. Journal of General Microbiology 70:351–360
     [Google Scholar]
  23. Höfer M., Becker J.-U., Brand K., Deckner K., Betz A. 1969; A study of the enzyme equipment of the yeast Rhodotorula gracilis. FEBS Letters 3:322–324
     [Google Scholar]
  24. Inoue H., Lowenstein J.M. 1975; Acetylcoenzyme A carboxylase from rat liver. Methods in Enzymology 35:3–11
     [Google Scholar]
  25. Kessell R.H.J. 1968; Fatty acids of Rhodotorula gracilis: fat production in submerged culture and the particular effect of pH value. Journal of Applied Bacteriology 31:220–231
     [Google Scholar]
  26. Knowles C.J. 1977; Microbial metabolic regulation by adenine nucleotide pools. Symposia of the Society for General Microbiology 27:241–283
     [Google Scholar]
  27. Kornberg A. 1955; Isocitric dehydrogenase of yeast. Methods in Enzymology 1:705–709
     [Google Scholar]
  28. Kornberg A., Pricer W.E. 1951; Di- and triphosphopyridine nucleotide isocitrate dehydrogenases in yeast. Journal of Biological Chemistry 189:123–136
     [Google Scholar]
  29. Lane M.D., Moss J., Polakis S.E. 1974; Acetyl-coenzyme A carboxylase. Current Topics in Cellular Regulation 8:139–195
     [Google Scholar]
  30. Lowenstein J.M. 1971; The pyruvate dehydrogenase complex and the citric acid cycle. In Comprehensive Biochemistry 18S pp. 1–55 Florkin M., Stotz E. H. Edited by Amsterdam, London & New York: Elsevier;
     [Google Scholar]
  31. Marchal R., Vandecasteele J.P., Metche M. 1977; Regulation of the central metabolism in relation to citric acid production in Saccharo mycopsis lipolytica. Archives of Microbiology 113:99–104
     [Google Scholar]
  32. Markall J.C., Lane M.D. 1977; Role of pyruvate carboxylase in fatty acid synthesis: alterations during preadipocyte differentiations. Biochemical and Biophysical Research Communications 79:720–725
     [Google Scholar]
  33. Matsuhashi M. 1969; Acetyl-coenzyme A carboxylase from yeast. Methods in Enzymology 14:3–8
     [Google Scholar]
  34. Miller A.L., Atkinson D.E. 1972; Response of yeast pyruvate carboxylase to the adenylate energy charge and other regulatory parameters. Archives of Biochemistry and Biophysics 152:531–538
     [Google Scholar]
  35. Mitsdshima K., Shinmyo A., Enatsu T. 1978; Control of citrate and 2-oxoglutarate formation in Candida lipolytica mitochondria by adenine nucleotides. Biochimica et biophysica acta 538:481–492
     [Google Scholar]
  36. Neijssel O.M., Hueting S., Tempest D.W. 1977; Glucose transport capacity is not the rate-limiting step in the growth of some wild-type strains of Escherichia coli and Klebsiella aerogenes. FEMS Microbiology Letters 2:1–3
     [Google Scholar]
  37. Rasmussen P.K., Klein H.P. 1967; Regulation of acetyl-CoA carboxylase of Saccharomyces cerevisiae. Biochemical and Biophysical Research Communications 28:415–419
     [Google Scholar]
  38. Rasmussen P.K., Klein H.P. 1968; Activation of fatty acid synthesis in cell-free extracts of Saccharomyces cerevisiae. Journal of Bacteriology 95:157–161
     [Google Scholar]
  39. Ratledge C. 1978; Lipids and fatty acids. In Economic Microbiology 2 pp. 263–301 Rose A. H. Edited by New York & London: Academic Press;
     [Google Scholar]
  40. 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]
  41. Rattray J.B.M., Schibeci A., Kidby D.K. 1975; Lipids of yeasts. Bacteriological Reviews 39:197–231
     [Google Scholar]
  42. Rouser G., Kritchevsky G., Yamamoto A. 1967; Column chromatographic and associated procedures for separation and determination of phosphatides and glycolipids. In Lipid Chromatographic Analysis pp. 99–162 Marsetti G. W. Edited by London: Edward Arnold;
     [Google Scholar]
  43. Serrano R., Gancedo J.M., Gancedo C. 1973; Assay of yeast enzymes in situ. A potential tool in regulation studies. European Journal of Biochemistry 34:479–482
     [Google Scholar]
  44. Simon E.J., Shemin D. 1953; Synthesis of acetyl-Co A. Journal of the American Chemical Society 75:2520
     [Google Scholar]
  45. Sols A., Gancedo C., Delafuente G. 1971; Energy-yielding metabolism in yeasts. In The Yeasts 2 pp. 271–308 Rose A. H. Edited by New York & London: Academic Press;
     [Google Scholar]
  46. Srere P.A. 1962; Citrate-cleavage enzyme. Methods in Enzymology 5:641–644
     [Google Scholar]
  47. Srere P.A. 1972; The citrate enzymes: their structures, mechanisms and biological functions. Current Topics in Cellular Regulation 5:229–283
     [Google Scholar]
  48. Strehler B.L., McElroy W.D. 1957; Assay of adenosine triphosphate. Methods in Enzymology 3:871–873
     [Google Scholar]
  49. Thomas K.C., Dawson P.S.S. 1977; Variations in theadenylate energy charge during phased growth (cell cycle) of Candida utilis under energy excess and energy-limiting growth conditions. Journal of Bacteriology 132:36–43
     [Google Scholar]
  50. Thorpe R.F., Ratledge C. 1972; Fatty acid distribution in triglycerides of yeasts grown on glucose or n-alkanes. Journal of General Microbiology 12:151–163
     [Google Scholar]
  51. Uzuka Y., Kanamori T., Koga T., Tanaka K., Nagauma T. 1975; Isolation and chemical composition of intracellular oil globules from the yeast Lipomyces starkeyi. Journal of General and Applied Microbiology 21:157–168
     [Google Scholar]
  52. Vagelos P.R. 1974; Biosynthesis of saturated fatty acids. In MTP International Review of Science: Biochemistry (Series One), Biochemistry of Lipids 4 pp. 99–140 Goodwin T. W. Edited by London: Butterworths;
     [Google Scholar]
  53. Volpe J.J., Vagelos P.R. 1976; Biosynthesis of fatty acids. Physiological Reviews 56:339–417
     [Google Scholar]
  54. Webster L.T. 1969; Acetyl-CoA carboxylase. Methods in Enzymology 13:375–381
     [Google Scholar]
  55. White D., Klein H.P. 1965; Factors affecting fatty acid synthesis in cell-free preparations from Saccharomyces cerevisiae. Biochemical and Biophysical Research Communications 20:78–84
     [Google Scholar]
  56. White D., Klein H.P. 1966; Effects of α-glycerophosphate and of palmitoyl-coenzyme A on lipid synthesis in yeast extracts. Journal of Bacteriology 91:1218–1223
     [Google Scholar]
  57. Whitworth D.A., Ratledge C. 1975; An analysis of intermediary metabolism and its control in a fat-synthesizing yeast (Candida 107) growing on glucose or alkanes. Journal of General Microbiology 88:275–288
     [Google Scholar]
  58. Whitworth D.A., Ratledge C. 1977; Phosphoketolase in Rhodotorula graminis and other yeasts. Journal of General Microbiology 102:397–401
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-114-2-361
Loading
A Biochemical Explanation for Lipid Accumulation in Candida 107 and Other Oleaginous Micro-organisms
Microbiology 114, 361 (1979); https://doi.org/10.1099/00221287-114-2-361
/content/journal/micro/10.1099/00221287-114-2-361
/content/journal/micro/10.1099/00221287-114-2-361
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