Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme
Acid phosphatase from the yeast Saccharomyces cerevisiae was purified to homogeneity as ascertained by ultracentrifugation and electrophoresis. The purification procedure involved mechanical cell disruption, ethanol precipitation, chromatography on DEAE-cellulose, gel filtration on Sepharose 4B. The...
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| Format: | Article |
| Language: | English |
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Polish Botanical Society
2015-01-01
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| Series: | Acta Societatis Botanicorum Poloniae |
| Online Access: | https://pbsociety.org.pl/journals/index.php/asbp/article/view/4911 |
| _version_ | 1818239191444619264 |
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| author | W. Wątorek B. Morawiecka |
| author_facet | W. Wątorek B. Morawiecka |
| author_sort | W. Wątorek |
| collection | DOAJ |
| description | Acid phosphatase from the yeast Saccharomyces cerevisiae was purified to homogeneity as ascertained by ultracentrifugation and electrophoresis. The purification procedure involved mechanical cell disruption, ethanol precipitation, chromatography on DEAE-cellulose, gel filtration on Sepharose 4B. The sedimentation constant S200.580 of the purified enzyme was 15.4 S. Carbohydrate content accounted for 50% of the total molecular weight of the enzyme. The optimum pH for purified enzyme was 3.0-3.5, it was stable at pH 3.0-5.0 at room temperature. After 10 min. incubation at 45° C, 50 per cent of the enzymatic activity was lost. Michaelis constant was found to be 1.3 x 10-4 M for p-nitrophenylphosphate and 5 x 10-4 M for 3-glycerophosphate as substrates. The enzyme was inhibited by Hg2+, Cu2+, Fe3+, molybdate, phosphate, arsenate, fluoride ions. Inhibition caused by fluoride ions was noncompetitive, by phosphate - competitive, 5 M urea inactivated the enzyme completely, inactivation was reversible at urea concentration below 2,5 M. |
| first_indexed | 2024-12-12T12:53:37Z |
| format | Article |
| id | doaj.art-fcd22aee6b64490598de5a5a748cb8df |
| institution | Directory Open Access Journal |
| issn | 2083-9480 |
| language | English |
| last_indexed | 2024-12-12T12:53:37Z |
| publishDate | 2015-01-01 |
| publisher | Polish Botanical Society |
| record_format | Article |
| series | Acta Societatis Botanicorum Poloniae |
| spelling | doaj.art-fcd22aee6b64490598de5a5a748cb8df2022-12-22T00:23:56ZengPolish Botanical SocietyActa Societatis Botanicorum Poloniae2083-94802015-01-0146217318610.5586/asbp.1977.0144114Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzymeW. Wątorek0B. Morawiecka1University of WrocławUniversity of WrocławAcid phosphatase from the yeast Saccharomyces cerevisiae was purified to homogeneity as ascertained by ultracentrifugation and electrophoresis. The purification procedure involved mechanical cell disruption, ethanol precipitation, chromatography on DEAE-cellulose, gel filtration on Sepharose 4B. The sedimentation constant S200.580 of the purified enzyme was 15.4 S. Carbohydrate content accounted for 50% of the total molecular weight of the enzyme. The optimum pH for purified enzyme was 3.0-3.5, it was stable at pH 3.0-5.0 at room temperature. After 10 min. incubation at 45° C, 50 per cent of the enzymatic activity was lost. Michaelis constant was found to be 1.3 x 10-4 M for p-nitrophenylphosphate and 5 x 10-4 M for 3-glycerophosphate as substrates. The enzyme was inhibited by Hg2+, Cu2+, Fe3+, molybdate, phosphate, arsenate, fluoride ions. Inhibition caused by fluoride ions was noncompetitive, by phosphate - competitive, 5 M urea inactivated the enzyme completely, inactivation was reversible at urea concentration below 2,5 M.https://pbsociety.org.pl/journals/index.php/asbp/article/view/4911 |
| spellingShingle | W. Wątorek B. Morawiecka Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme Acta Societatis Botanicorum Poloniae |
| title | Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme |
| title_full | Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme |
| title_fullStr | Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme |
| title_full_unstemmed | Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme |
| title_short | Acid phosphatase of the yeast Saccharomyces cerevisiae. Purification and properties of the enzyme |
| title_sort | acid phosphatase of the yeast saccharomyces cerevisiae purification and properties of the enzyme |
| url | https://pbsociety.org.pl/journals/index.php/asbp/article/view/4911 |
| work_keys_str_mv | AT wwatorek acidphosphataseoftheyeastsaccharomycescerevisiaepurificationandpropertiesoftheenzyme AT bmorawiecka acidphosphataseoftheyeastsaccharomycescerevisiaepurificationandpropertiesoftheenzyme |