1887

Abstract

Fractionated extracts of were found to contain the readily dissociable type of fatty acid synthase complex. For optimal activity the partially purified synthase required the presence of acetyl-CoA, malonyl-CoA, NADH and NADPH as well as a cell-free fraction containing the acyl carrier protein (ACP). When ACP-containing fractions obtained from grown in the presence of [1-C]pantothenate were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis, the radioactivity was located in a peak corresponding to a molecular weight of approx. 8500–9000. The biological activity of the ACP was relatively heat-stable and trypsin-resistant. When the labelled ACP-containing fraction was subjected to gel filtration on a calibrated Sephadex G-75 column, a radioactive peak corresponding to a molecular weight of 22000 was obtained that could participate in the CO-malonyl-CoA exchange reaction in the presence of the other requisite components.

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1981-11-01
2024-12-12
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References

  1. Anker H. S. 1970; A solubilizable acrylamide gel for electrophoresis. FEBS Letters 7:293
    [Google Scholar]
  2. Birge C. H., Vagelos P. R. 1972; Acyl carrier protein. Journal of Biological Chemistry 247:4921–4929
    [Google Scholar]
  3. Bloch K., Vance D. 1977; Control mechanisms in the synthesis of saturated fatty acids. Annual Review of Biochemistry 46:263–298
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  5. Brindley D. N., Matsumara S., Bloch K. 1969; Mycobacterium phlei fatty acid synthetase - a bacterial multienzyme complex. Nature; London: 224666–669
    [Google Scholar]
  6. Delwiche E. A. 1949; Vitamin requirement of the genus Propionibacterium. Journal of Bacteriology 58:395–398
    [Google Scholar]
  7. Fricke U. 1975; Tritosol: a new scintillation cocktail based on Triton X-100. Analytical Biochemistry 63:555–558
    [Google Scholar]
  8. Kawaguchi A., Okuda S. 1977; Fatty acid synthetase from Brevibacterium ammoniagenes: formation of monounsaturated fatty acids by a multienzyme complex. Proceedings of the National Academy of Sciences of the United States of America 74:3180–3183
    [Google Scholar]
  9. Layne E. 1957; Spectrophotometric and turbidimetric methods for measuring proteins. Methods in Enzymology 3:447–454
    [Google Scholar]
  10. Lynen F. 1961; Biosynthesis of saturated fatty acids. Federation Proceedings 20:941–951
    [Google Scholar]
  11. Majerus P. W. 1967; Acyl carrier protein. Structural requirements for function in fatty acid biosynthesis. Journal of Biological Chemistry 242:2325–2332
    [Google Scholar]
  12. Majerus P. W. 1968; Acyl carrier protein: effect of acetylation and tryptic hydrolysis on function in fatty acid synthesis. Science 159:428–430
    [Google Scholar]
  13. Majerus P. W., Alberts A. W., Vagelos P. R. 1969; Acyl carrier protein from Escherichia coli. Methods in Enzymology 14:43–50
    [Google Scholar]
  14. Matsumara S., Brindley D. N., Bloch K. 1970; Acyl carrier protein from Mycobacterium phlei. Biochemical and Biophysical Research Communications 38:369–377
    [Google Scholar]
  15. Overath P., Stumpf P. K. 1964; Fat metabolism in higher plants - properties of a soluble fatty acid synthetase from avocado mesocarp. Journal of Biological Chemistry 239:4103–4110
    [Google Scholar]
  16. Rossi A., Corcoran J. W. 1973; Identification of a multienzyme complex synthesizing fatty acids in the actinomycete Streptomyces erythreus. Biochemical and Biophysical Research Communications 50:597–602
    [Google Scholar]
  17. Simoni R. D., Criddle R. S., Stumpf P. K. 1967; Fat metabolism in higher plants. Purification and properties of plant and bacterial acyl carrier proteins. Journal of Biological Chemistry 242:573–581
    [Google Scholar]
  18. Stoops J. K., Arslanian M. J., Oh Y. H., Aune K. C., Vanaman T. C., Wakil S. J. 1975; Presence of two polypeptide chains comprising fatty acid synthetase. Proceedings of the National Academy of Sciences of the United States of America 72:1940–1944
    [Google Scholar]
  19. Vagelos P. R. 1974; Biosynthesis of saturated fatty acids. In Biochemistry of Lipids 4 pp. 99–140 Goodwin T. W. Edited by London, Baltimore & Tokyo: University Park Press.;
    [Google Scholar]
  20. Vanaman T. C., Wakil S. J., Hill R. L. 1968; The complete amino acid sequence of the acyl carrier protein of Escherichia coli. Journal of Biological Chemistry 243:6420–6431
    [Google Scholar]
  21. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  22. White H. B., Mitsuhashi O., Bloch K. 1971; Pyridine nucleotide requirements of fatty acid synthetases. Journal of Biological Chemistry 246:4751–4754
    [Google Scholar]
  23. Wood H. G., Jacobson B., Gerwin B. I., Northrop D. 1969; Oxaloacetate transcarboxylase from Propionibacterium. Methods in Enzymology 13:215–230
    [Google Scholar]
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