Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1976 Mar;125(3):999–1004. doi: 10.1128/jb.125.3.999-1004.1976

Utilization of D-asparagine by Saccharomyces cerevisiae.

P C Dunlop, R J Roon, H L Even
PMCID: PMC236176  PMID: 767332

Abstract

Yeast strains sigma1278b and Harden and Young, which synthesize only an internal constitutive form of L-asparaginase, do not grow on D-asparagine, as a sole source of nitrogen, and whole cell suspensions of these strains do not hydrolyze D-asparagine. Strains X2180-A2 and D273-10B, which possess an externally active form of asparaginase, are able to grow slowly on D-asparagine, and nitrogen-starved suspensions of these strains exhibit high activity toward the D-isomer. Nitrogen starvation of strain X218O-A2 results in coordinate increase of D- and L-asparaginase activity; the specific activity observed for the D-isomer is approximately 20% greater than that observed for the L-isomer. It was observed, in studies with cell extracts, that hydrolysis of D-asparagine occurred only with extracts from nitrogen-starved cells of strains that synthesize the external form of asparaginase. Furthermore, the activity of the extracts toward the D-isomer was always higher than that observed with the L-isomer. A 400-fold purified preparation of external asparaginase from Saccharomyces cerevisiae X218U-A2 hydrolyzed D-asparagine with an apparent Km of 0.23 mM and a Vmax of 38.7 mumol/min per mg of protein. D-Asparagine was a competitive inhibitor of L-asparagine hydrolysis and the Ki determined for this inhibition was approximately equal to its Km. These data suggest that D-asparagine is a good substrate for the external yeast asparaginase but is a poor substrate for the internal enzyme.

Full text

PDF
1001

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ballou C. E. Some aspects of the structure, immunochemistry, and genetic control of yeast mannans. Adv Enzymol Relat Areas Mol Biol. 1974;40(0):239–270. doi: 10.1002/9780470122853.ch6. [DOI] [PubMed] [Google Scholar]
  2. Broome J. D. Antilymphoma activity of L-asparaginase in vivo: clearance rates of enzyme preparations from guinea pig serum and yeast in relation to their effect on tumor growth. J Natl Cancer Inst. 1965 Dec;35(6):967–974. [PubMed] [Google Scholar]
  3. Campbell H. A., Mashburn L. T. L-Asparaginase EC-2 from Escherichia coli. Some substrate specificity characteristics. Biochemistry. 1969 Sep;8(9):3768–3775. doi: 10.1021/bi00837a042. [DOI] [PubMed] [Google Scholar]
  4. Cooney D. A., Handschumacher R. E. L-asparaginase and L-asparagine metabolism. Annu Rev Pharmacol. 1970;10:421–440. doi: 10.1146/annurev.pa.10.040170.002225. [DOI] [PubMed] [Google Scholar]
  5. Dunlop P. C., Roon R. J. L-Asparaginase of Saccharomyces cerevisiae: an extracellular Enzyme. J Bacteriol. 1975 Jun;122(3):1017–1024. doi: 10.1128/jb.122.3.1017-1024.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gascón S., Neumann N. P., Lampen J. O. Comparative study of the properties of the purified internal and external invertases from yeast. J Biol Chem. 1968 Apr 10;243(7):1573–1577. [PubMed] [Google Scholar]
  7. Howard J. B., Carpenter F. H. L-asparaginase from Erwinia carotovora. Substrate specificity and enzymatic properties. J Biol Chem. 1972 Feb 25;247(4):1020–1030. [PubMed] [Google Scholar]
  8. Joiris C. R., Grenson M. Spécificité et régulation d'une perméase des acis aminés dicarboxyliques chez "Saccharomyces crevisiae". Arch Int Physiol Biochim. 1969 Feb;77(1):154–156. [PubMed] [Google Scholar]
  9. LaRue T. A., Spencer J. F. The utilization of D-amino acids by yeasts. Can J Microbiol. 1967 Jul;13(7):777–788. doi: 10.1139/m67-103. [DOI] [PubMed] [Google Scholar]
  10. Roon R. J., Even H. L., Dunlop P., Larimore F. L. Methylamine and ammonia transport in Saccharomyces cerevisiae. J Bacteriol. 1975 May;122(2):502–509. doi: 10.1128/jb.122.2.502-509.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Rytka J. Positive selection of general amino acid permease mutants in Saccharomyces cerevisiae. J Bacteriol. 1975 Feb;121(2):562–570. doi: 10.1128/jb.121.2.562-570.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Smith W. L., Ballou C. E. Immunochemical characterization of the mannan component of the external invertase (beta-fructofuranosidase) of Saccharomyces cerevisiae. Biochemistry. 1974 Jan 15;13(2):355–361. doi: 10.1021/bi00699a021. [DOI] [PubMed] [Google Scholar]
  13. Smith W. L., Ballou C. E. The effect of dithiothreitol on external yeast invertase. Biochem Biophys Res Commun. 1974 Jul 10;59(1):314–321. doi: 10.1016/s0006-291x(74)80208-1. [DOI] [PubMed] [Google Scholar]
  14. To-E A., Ueda Y., Kakimoto S. I., Oshima Y. Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J Bacteriol. 1973 Feb;113(2):727–738. doi: 10.1128/jb.113.2.727-738.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ueda Y., To-E A., Oshima Y. Isolation and characterization of recessive, constitutive mutations for repressible acid phosphatase synthesis in Saccharomyces cerevisiae. J Bacteriol. 1975 Jun;122(3):911–922. doi: 10.1128/jb.122.3.911-922.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES