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THE INFLUENCE OF AMINO ACIDS ON THE REACTIVATION OF YEAST INVERTASE.

H Wagreich1, W Halpert, A Hirschman

  • 1Pediatric Research Laboratories, The Jewish Hospital of Brooklyn, New York, and the Department of Chemistry, the College of the City of New York.

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Sulfur-containing compounds like cysteine can reactivate acid-inactivated yeast invertase. However, oxidized forms and certain substitutions inhibit this enzyme reactivation process.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Protein Chemistry

Background:

  • Yeast invertase is crucial for sucrose hydrolysis.
  • Enzyme activity can be modulated by various chemical agents.
  • Understanding enzyme reactivation mechanisms is vital for biotechnology.

Purpose of the Study:

  • To investigate the effect of amino acids and sulfur-containing compounds on acid-inactivated yeast invertase reactivation.
  • To identify specific chemical groups responsible for modulating enzyme recovery.
  • To elucidate the mechanism of enzyme reactivation by thiol compounds.

Main Methods:

  • Enzyme inactivation using acidic conditions.
  • Incubation of inactivated yeast invertase with various amino acids and sulfur-containing compounds.
  • Spectrophotometric assay to measure invertase activity and reactivation rates.
  • Testing the effect of substitutions on the thiol group.

Main Results:

  • 16 amino acids lacking thiol or disulfide groups did not influence reactivation.
  • Compounds with thiol groups (cysteine, glutathione, homocysteine, thiophenol, thioglycolic acid) accelerated reactivation.
  • Oxidized sulfur compounds (cystine, oxidized glutathione, homocystine) inhibited reactivation.
  • Native invertase activity was unaffected by these compounds.
  • Substitution of the hydrogen in homocysteine's thiol group abolished acceleration.
  • Increased contact time with cysteine enhanced reactivation velocity.
  • Cysteine's acceleration effect was inversely proportional to the control enzyme rate.

Conclusions:

  • Thiol and disulfide groups play a critical role in yeast invertase reactivation.
  • Reduced sulfur compounds promote enzyme recovery, while oxidized forms inhibit it.
  • The SH group's availability is essential for the accelerative effect on enzyme reactivation.