Asparaginase II of Saccharomyces cerevisiae: Comparison of Enzyme Stability in Vivo and in Vitro

Kyu Won Kim, Robert J Roon

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Asparaginase II of Saccharomyces cerevisiae is a cell wall mannan containing glycoprotein. Recent studies have demonstrated that asparaginase II activity increases in exponentially growing cell cultures and then decreases as the cells enter the stationary phase. Enzyme inactivation has been attributed to a Zn2+-dependent protease which is synthesized de novo during the late exponential phase [Pauling, K. D., & Jones, G. E. (1980) J. Gen. Microbiol. 117, 423–430; Pauling, K. D., & Jones, G. E. (1980) Biochim. Biophys. Acta 616, 271–282]. We have investigated the mechanism of asparaginase II inactivation using both whole cell suspensions and highly purified enzyme. Our data indicate that the rate of asparaginase II inactivation in cell suspensions is primarily influenced by pH changes that occur as a consequence of cell growth and glucose fermentation and that enzyme inactivation is not dependent on Zn2+ or on de novo protein synthesis. Also, in vitro studies with purified enzyme show kinetics of inactivation that are similar to those observed in vivo. Consequently, involvement of a yeast protease in the inactivation process is relatively unlikely.

Original languageEnglish (US)
Pages (from-to)2704-2707
Number of pages4
JournalBiochemistry
Volume22
Issue number11
DOIs
StatePublished - Jan 1 1983

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Enzyme Stability
Asparaginase
Yeast
Saccharomyces cerevisiae
Enzymes
Suspensions
Peptide Hydrolases
Mannans
Enzyme kinetics
Cell growth
Cell culture
Fermentation
Cell Wall
Glycoproteins
Cell Culture Techniques
Yeasts
Cells
Glucose
In Vitro Techniques
Growth

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Asparaginase II of Saccharomyces cerevisiae : Comparison of Enzyme Stability in Vivo and in Vitro. / Kim, Kyu Won; Roon, Robert J.

In: Biochemistry, Vol. 22, No. 11, 01.01.1983, p. 2704-2707.

Research output: Contribution to journalArticle

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abstract = "Asparaginase II of Saccharomyces cerevisiae is a cell wall mannan containing glycoprotein. Recent studies have demonstrated that asparaginase II activity increases in exponentially growing cell cultures and then decreases as the cells enter the stationary phase. Enzyme inactivation has been attributed to a Zn2+-dependent protease which is synthesized de novo during the late exponential phase [Pauling, K. D., & Jones, G. E. (1980) J. Gen. Microbiol. 117, 423–430; Pauling, K. D., & Jones, G. E. (1980) Biochim. Biophys. Acta 616, 271–282]. We have investigated the mechanism of asparaginase II inactivation using both whole cell suspensions and highly purified enzyme. Our data indicate that the rate of asparaginase II inactivation in cell suspensions is primarily influenced by pH changes that occur as a consequence of cell growth and glucose fermentation and that enzyme inactivation is not dependent on Zn2+ or on de novo protein synthesis. Also, in vitro studies with purified enzyme show kinetics of inactivation that are similar to those observed in vivo. Consequently, involvement of a yeast protease in the inactivation process is relatively unlikely.",
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N2 - Asparaginase II of Saccharomyces cerevisiae is a cell wall mannan containing glycoprotein. Recent studies have demonstrated that asparaginase II activity increases in exponentially growing cell cultures and then decreases as the cells enter the stationary phase. Enzyme inactivation has been attributed to a Zn2+-dependent protease which is synthesized de novo during the late exponential phase [Pauling, K. D., & Jones, G. E. (1980) J. Gen. Microbiol. 117, 423–430; Pauling, K. D., & Jones, G. E. (1980) Biochim. Biophys. Acta 616, 271–282]. We have investigated the mechanism of asparaginase II inactivation using both whole cell suspensions and highly purified enzyme. Our data indicate that the rate of asparaginase II inactivation in cell suspensions is primarily influenced by pH changes that occur as a consequence of cell growth and glucose fermentation and that enzyme inactivation is not dependent on Zn2+ or on de novo protein synthesis. Also, in vitro studies with purified enzyme show kinetics of inactivation that are similar to those observed in vivo. Consequently, involvement of a yeast protease in the inactivation process is relatively unlikely.

AB - Asparaginase II of Saccharomyces cerevisiae is a cell wall mannan containing glycoprotein. Recent studies have demonstrated that asparaginase II activity increases in exponentially growing cell cultures and then decreases as the cells enter the stationary phase. Enzyme inactivation has been attributed to a Zn2+-dependent protease which is synthesized de novo during the late exponential phase [Pauling, K. D., & Jones, G. E. (1980) J. Gen. Microbiol. 117, 423–430; Pauling, K. D., & Jones, G. E. (1980) Biochim. Biophys. Acta 616, 271–282]. We have investigated the mechanism of asparaginase II inactivation using both whole cell suspensions and highly purified enzyme. Our data indicate that the rate of asparaginase II inactivation in cell suspensions is primarily influenced by pH changes that occur as a consequence of cell growth and glucose fermentation and that enzyme inactivation is not dependent on Zn2+ or on de novo protein synthesis. Also, in vitro studies with purified enzyme show kinetics of inactivation that are similar to those observed in vivo. Consequently, involvement of a yeast protease in the inactivation process is relatively unlikely.

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