Stabilization and purification of the secondary metabolism specific enzyme, m-hydroxybenzylalcohol dehydrogenase
| Autor: | Richard E. Scott, K. S. Lam, G. M. Gaucher |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Immunology
Dehydrogenase Buffers Chemical Fractionation Applied Microbiology and Biotechnology Microbiology Chromatography Affinity Protein purification Genetics Penicillium urticae Secondary metabolism Molecular Biology Specific enzyme chemistry.chemical_classification Chromatography biology Penicillium General Medicine Fungi imperfecti Chromatography Ion Exchange biology.organism_classification Alcohol Oxidoreductases Patulin Enzyme Biochemistry chemistry Stationary phase Chromatography Gel Electrophoresis Polyacrylamide Gel Half-Life |
| Zdroj: | Canadian Journal of Microbiology. 32:167-175 |
| ISSN: | 1480-3275 0008-4166 |
| DOI: | 10.1139/m86-033 |
| Popis: | m-Hydroxybenzylalcohol dehydrogenase (EC 1.1.1.97), a secondary metabolism associated protein from stationary phase cultures of Penicillium urticae, was stabilized in crude extracts prior to purification. Stabilization studies resulted in the formulation of an optimal cell breakage and purification buffer. This buffer increased the enzyme's in vitro half-life at 30 °C from 14 to over 800 min which greatly aided purification and enhanced yields. Purification was achieved by salt fractionation, size-exclusion chromatography, affinity chromatography, and ion-exchange chromatography. The 1200-fold purified protein gave only one major band by sodium dodecyl sulphate – polyacrylamide gel electrophoresis. |
| Databáze: | OpenAIRE |
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