Fast purification and kinetic studies of the glycerol-3-phosphate dehydrogenase from the yeast Saccharomyces cerevisiae
| Autor: | Manfred Rizzi, Jingmin Cai, Uwe Theobald, Markus Pietzsch |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Glycerol
Gel electrophoresis Chromatography biology Chemistry Saccharomyces cerevisiae Ion chromatography Glycerolphosphate Dehydrogenase Bioengineering Dehydrogenase General Medicine NAD biology.organism_classification Binding Competitive Applied Microbiology and Biotechnology Substrate Specificity Kinetics Glucose Glycerol-3-phosphate dehydrogenase Non-competitive inhibition Biochemistry Product inhibition Enzyme kinetics Biotechnology |
| Zdroj: | Journal of Biotechnology. 49:19-27 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/0168-1656(96)01509-x |
| Popis: | The glycerol-3-phosphate dehydrogenase has been purified from Saccharomyces cerevisiae 140-fold to electrophoretic homogeneity by a simple procedure involving affinity and ion exchange chromatography. The purified enzyme was most active at pH 6.8 and 51 degrees C. Its molecular mass was determined to be 45000 +/- 2000 Da by SDS-polyacrylamide gel electrophoresis. At physiological pH values the thermodynamic equilibrium constant was determined to be 3.5 x 10(-3) (M-1). Product inhibition as well as competitive inhibition patterns were found which clearly indicate that the kinetic mechanism of the glycerol-3-phosphate dehydrogenase is random bi-bi with the formation of dead-end complexes. In vivo concentrations of selected metabolites and kinetic expression for G3P-DH were used to explain regulatory properties of this enzyme under conditions of short-term glucose effect in Saccharomyces cerevisiae. |
| Databáze: | OpenAIRE |
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