Molecular and General Genetics MGG

, Volume 139, Issue 4, pp 303–309 | Cite as

Citrate synthaseless glutamic acid auxotroph of Saccharomyces cerevisiae

  • John P. Burand
  • Robert Drillien
  • J. K. Bhattacharjee
Article
Received:

Summary

Relationship of citrate synthase (EC 4.1.3.7) to the biosynthesis of glutamic acid was investigated by characterizing a new glutamic acid auxotroph FL100-D1 (glu 3) of Saccharomyces cerevisiae. Nutritional requirement of the mutant was satisfied by L-glutamic acid, L-glutamic acid peptide as well as several analogs of glutamic acid, but not by proline, ornithine, arginine, lysine or aspartic acid. The mutant was unable to utilize nonfermentable carbon sources, glycerol, acetate or lactate. Mutant glu 3 unlike aconitaseless glutamic acid auxotroph glu 1, failed to accumulate 14C-citric acid in vivo from 1-14C-sodium acetate or U-14C-glutamic acid. Both spectrophotometric and radioactive assay procedures demonstrated a lack of significant citrate synthase activity in the dialysed extract of the mutant compared to the wild type strain. Mutant glu 3 complemented with glu 1 and glu 2 individually in vivo and exhibited a significant aconitase (EC 4.2.1.3) activity in vitro.

Keywords

Proline Lysine Arginine Glutamic Acid Saccharomyces Cerevisiae 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. Battacharjee, J.K.: A glucose EMB agar method for detection of respiratory deficiency in yeast. Canad. J. Microbiol. 13, 1116–1117 (1967)Google Scholar
  2. Bhattacharjee, J.K., Strassman, M.: Accumulation of tricarboxylic acid related to lysine biosynthesis in a yeast mutant. J. biol. Chem. 242, 2542–2546 (1967)Google Scholar
  3. Carls, R.A., Hanson, R.S.: Isolation and characterization of tricarboxylic acid cycle mutants of Bacillus subtilis. J. Bact. 106, 848–855 (1971)Google Scholar
  4. Coleman, J.S., Bhattacharjee, J.K.: Regulation of citrate synthase activity of Saccharomyces cerevisiae. Antonie Van Leeuwenhoek 41, 53–60 (1975)Google Scholar
  5. Crocker, Jr., W.H., Bhattacharjiee, J.K.: Glutamic acid biosynthesis in Saccaromyces: Accumulation of tricarboxylic acid cycle intermediates in a glutamate auxotroph. Appl. Microbiol. 26, 303–308 (1973)Google Scholar
  6. Drillien, R., Lacroute, F.: Ureido-succinic acid uptake in yeast and some aspects of its regulation. J. Bact. 109, 203–208 (1972)Google Scholar
  7. Flechtner, V.R., Hanson, R.S.: Regulation of the tricarboxylic acid cycle in bacteria: a comparison of citrate synthesis from different bacteria. Biochim. biophys. Acta (Amst.) 222, 256–264 (1970)Google Scholar
  8. Fortnagel, P.: The regulation of aconitase and isocitrate dehydrogenase in sporulation mutants of Bacillus subtilis. Biochim. biophys. Acta (Amst.) 222, 290–298 (1970)Google Scholar
  9. Gilvarg, C., Davis, B.D.: The role of the tricarboxylic acid cycle in acetate oxidation in Escherichia coli. J. biol. Chem. 222, 307–319 (1956)Google Scholar
  10. Gornall, A.G., Bardawill, C.J., David, M.M.: Determination of serum protein by means of the biuret reaction. J. biol. Chem. 177, 751–766 (1949)Google Scholar
  11. Lacroute, F.: Non-mendelian mutation allowing ureidosuccinic acid uptake in yeast. J. Bact. 106, 519–522 (1971)Google Scholar
  12. Ogur, M., Coker, L., Ogur, S.: Glutamate auxotroph in Saccharomyces. The biochemical lesion in the glt 1 mutants. Biochem. biophys. Res. Commun. 14, 193–197 (1964)Google Scholar
  13. Ogur, M., Roshanmanesh, A., Ogur, S.: Tricarboxylic acid cycle mutants in Saccharomyces: Comparison of independently derived mutants. Science 147, 1590 (1965)Google Scholar
  14. Ohne, M.: Regulation of aconitase synthesis in Bacillus subtilis: induction feedback repression and catabolic repression. J. Bact. 117, 1295–1305 (1974)Google Scholar
  15. Parvin, R.: Citrate synthase from yeast, p. 16–19. In: Methods in Enzymology, Vol. 13, (S.O. Colowick and N.O. Kaplan eds.). New York: Academic Press Inc. 1969Google Scholar
  16. Racker, E.: Spectrophotometric measurements of the enzymatic formation of fumaric acid and cis-aconitic acids. Biochim. biophys. Acta (Amst.) 4, 211–214 (1950)Google Scholar
  17. Sinha, A.K., Bhattacharjee, J.K.: Control of a lysine biosynthetic step by two unliked genes of Saccharomyces. Biochem. biophys. Res. Commun. 39, 1205–1209 (1970)Google Scholar
  18. Strassman, M., Ceci, L.N.: Enzymatic formation of cis-homoaconitic acid an intermediate in lysine biosynthesis in yeast. J. biol. Chem. 241, 5401–5407 (1966)Google Scholar
  19. Wieland, O., Weiss, L., Eger-Neufeldt, I.: Inhibition of enzymatic citric acid synthesis by long-chained acetyl-thioesters of coenzyme A. Biochem. Z. 339, 501–513 (1964)Google Scholar
  20. Wright, J.A., Moeba, P., Sanwal, B.D.: Allosteric regulation of the citrate synthase of Escherichia coli by α-ketoglutarate. Biochem. biophys. Res. Commun. 29, 34–38 (1967)Google Scholar

Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • John P. Burand
    • 1
  • Robert Drillien
    • 2
  • J. K. Bhattacharjee
    • 1
  1. 1.Department of MicrobiologyMiami UniversityOxford
  2. 2.Laboratoire de Génétique PhysiologiqueInstitut de BontaniqueStrasbourgFrance

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