Utilization of D asparagine by Saccharomyces cerevisiae

P. C. Dunlop, Robert J Roon, H. L. Even

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Abstract

Yeast strains sigma 1278b 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 X2180 A2 results in coordinate increase of D- and L asparaginase activity; the specific activity observed for the D isomer is 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 S. cerevisiae X2180 A2 hydrolyzed D asparagine with and apparent K(m) of 0.23 mM and a V(max) of 38.7 μmol/min per mg of protein. D Asparagine was a competitive inhibitor of L asparagine hydrolysis and the K(i) determined for this inhibition was approximately equal to its K(m). These data suggest the D asparagine is a good substrate for the external yeast asparaginase but is a poor substrate for the internal enzyme.

Original languageEnglish (US)
Pages (from-to)999-1004
Number of pages6
JournalJournal of Bacteriology
Volume125
Issue number3
StatePublished - Dec 1 1976

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Asparagine
Asparaginase
Saccharomyces cerevisiae
Nitrogen
varespladib methyl
Suspensions
Hydrolysis
Yeasts
L Forms
Starvation
Cell Extracts
Enzymes

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Utilization of D asparagine by Saccharomyces cerevisiae. / Dunlop, P. C.; Roon, Robert J; Even, H. L.

In: Journal of Bacteriology, Vol. 125, No. 3, 01.12.1976, p. 999-1004.

Research output: Contribution to journalArticle

Dunlop, PC, Roon, RJ & Even, HL 1976, 'Utilization of D asparagine by Saccharomyces cerevisiae', Journal of Bacteriology, vol. 125, no. 3, pp. 999-1004.
Dunlop, P. C. ; Roon, Robert J ; Even, H. L. / Utilization of D asparagine by Saccharomyces cerevisiae. In: Journal of Bacteriology. 1976 ; Vol. 125, No. 3. pp. 999-1004.
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abstract = "Yeast strains sigma 1278b 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 X2180 A2 results in coordinate increase of D- and L asparaginase activity; the specific activity observed for the D isomer is 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 S. cerevisiae X2180 A2 hydrolyzed D asparagine with and apparent K(m) of 0.23 mM and a V(max) of 38.7 μmol/min per mg of protein. D Asparagine was a competitive inhibitor of L asparagine hydrolysis and the K(i) determined for this inhibition was approximately equal to its K(m). These data suggest the D asparagine is a good substrate for the external yeast asparaginase but is a poor substrate for the internal enzyme.",
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N2 - Yeast strains sigma 1278b 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 X2180 A2 results in coordinate increase of D- and L asparaginase activity; the specific activity observed for the D isomer is 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 S. cerevisiae X2180 A2 hydrolyzed D asparagine with and apparent K(m) of 0.23 mM and a V(max) of 38.7 μmol/min per mg of protein. D Asparagine was a competitive inhibitor of L asparagine hydrolysis and the K(i) determined for this inhibition was approximately equal to its K(m). These data suggest the D asparagine is a good substrate for the external yeast asparaginase but is a poor substrate for the internal enzyme.

AB - Yeast strains sigma 1278b 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 X2180 A2 results in coordinate increase of D- and L asparaginase activity; the specific activity observed for the D isomer is 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 S. cerevisiae X2180 A2 hydrolyzed D asparagine with and apparent K(m) of 0.23 mM and a V(max) of 38.7 μmol/min per mg of protein. D Asparagine was a competitive inhibitor of L asparagine hydrolysis and the K(i) determined for this inhibition was approximately equal to its K(m). These data suggest the D asparagine is a good substrate for the external yeast asparaginase but is a poor substrate for the internal enzyme.

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