Purification and characterisation of a metallopeptidase of Candida albicans
| Autor: | Machhour Ghazali, J. L. Jacquemin, Marie-Hélène Rodier, B E el Moudni, Christine Barrault |
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| Rok vydání: | 1995 |
| Předmět: |
Microbiology (medical)
Metallopeptidase Aminopeptidases Microbiology Michaelis–Menten kinetics High-performance liquid chromatography Aminopeptidase Substrate Specificity Candida albicans Humans Protease Inhibitors Isoelectric Point Chromatography High Pressure Liquid Edetic Acid Candida chemistry.chemical_classification biology Metalloendopeptidases General Medicine Hydrogen-Ion Concentration Chromatography Ion Exchange biology.organism_classification Yeast Corpus albicans Enzyme Biochemistry chemistry Chromatography Gel Electrophoresis Polyacrylamide Gel Phenanthrolines |
| Zdroj: | Journal of Medical Microbiology. 43:282-288 |
| ISSN: | 1473-5644 0022-2615 |
| Popis: | A novel aminopeptidase was purified by high performance liquid chromatography from a cytosoluble 100,000 g extract of Candida albicans on the basis of its ability to cleave L-arginine 7-amino-4-methylcoumarin. The purification factor was 36 and the yield was 20%. The native enzyme had a mol. wt of 52 kDa as demonstrated by SDS-PAGE in the presence or absence of reducing conditions and exhibited an iso-electric point of 4.3. The aminopeptidase showed optimum activity at pH 7.2, a Michaelis constant of c. 50 microM and a Vmax at 19 mM AMC released/min/mg of protein for L-Arg-AMC. This enzyme was shown to cleave at low affinity L-leucine-7-amino-4-methylcoumarin as demonstrated by the spectrofluorimetric method. The enzyme was strongly inhibited by specific metallo-enzyme inhibitors-EDTA and o-phenanthroline. Furthermore, there is evidence that a similar or identical enzyme occurs in other C. albicans clinical isolates and other Candida spp. |
| Databáze: | OpenAIRE |
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