Purification and Characterization of an Extracellular Alkaline Phosphatase fromPenicillium Chrysogenum
| العنوان: | Purification and Characterization of an Extracellular Alkaline Phosphatase fromPenicillium Chrysogenum |
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| المؤلفون: | Michael Politino, Joanne Brown, John J. Usher |
| المصدر: | Preparative Biochemistry and Biotechnology. 26:171-181 |
| بيانات النشر: | Informa UK Limited, 1996. |
| سنة النشر: | 1996 |
| مصطلحات موضوعية: | Phosphatase, Etoposide Phosphate, Penicillium chrysogenum, Biochemistry, chemistry.chemical_compound, Endoglycosidase H, Organophosphorus Compounds, Magnesium, Chromatography, High Pressure Liquid, Etoposide, chemistry.chemical_classification, Manganese, Chromatography, biology, Chemistry, General Medicine, Alkaline Phosphatase, Chromatography, Ion Exchange, biology.organism_classification, Phosphate, Enzyme assay, Molecular Weight, Kinetics, Zinc, Enzyme, Chromatography, Gel, biology.protein, Alkaline phosphatase, Electrophoresis, Polyacrylamide Gel, Biotechnology |
| الوصف: | An extracellular alkaline phosphatase from Penicillium chrysogenum was purified to homogeneity using DEAE ion-exchange chromatography and size exclusion chromatography. SDS-PAGE of the purified enzyme indicated a molecular weight of 58,000. The mobility of the native enzyme on a Superose 12 column suggests that the active form of the enzyme is a monomer. The enzyme catalyzes the hydrolysis of phosphate from a variety of substrates including p-nitrophenyl phosphate, alpha-naphthyl phosphate and the anti-tumor compound etoposide phosphate. The apparent K(m) for the substrate p-nitrophenyl phosphate is 1.3 mM and the enzyme is inhibited by inorganic phosphate. The pH optimum of the enzyme is 9.0 with a broad optimal temperature range between 40 and 50 degrees C. The isoelectric point of the enzyme is approximately 5.5. The enzyme is a glycoprotein; digestion with endoglycosidase H indicates that the protein consists primarily of N-linked carbohydrates. Enzymatic activity is enhanced by the addition of divalent cations such as Mg+2 and Mn+2 and inhibited by addition of a chelator such as EDTA suggesting a metal ion requirement. The enzyme was found to be an inexpensive catalyst for the conversion of etoposide phosphate to etoposide in the manufacture of this anti-tumor compound. |
| تدمد: | 1532-2297 1082-6068 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a48b45b2974bd677af6f684f9eb8a04c https://doi.org/10.1080/10826069608000063 |
| رقم الأكسشن: | edsair.doi.dedup.....a48b45b2974bd677af6f684f9eb8a04c |
| قاعدة البيانات: | OpenAIRE |
PDF full text from Publisher | تدمد: | 15322297 10826068 |
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