Purification and biochemical characterization of a novel protease from Penicillium digitatum - Use in bioactive peptides production
| Autor: | Neyssene, Aissaoui, Ferid, Abidi, Safa, Mahat, M Nejib, Marzouki |
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| Rok vydání: | 2014 |
| Předmět: |
Hydrolysis
Penicillium Temperature Muscle Proteins Hydrogen-Ion Concentration Chromatography Ion Exchange Perciformes Molecular Weight Phenylmethylsulfonyl Fluoride Surface-Active Agents Metals Enzyme Stability Chromatography Gel Animals Chemical Precipitation Electrophoresis Polyacrylamide Gel Protease Inhibitors Salts Protein Multimerization Serine Proteases Peptides Chromatography High Pressure Liquid |
| Zdroj: | Journal of basic microbiology. 54 |
| ISSN: | 1521-4028 |
| Popis: | This work reports the production of a novel serine protease enzyme (P. dig-protease) from the fungus Penicillium digitatum. The protease was purified from the culture supernatant to homogeneity using ammonium sulfate precipitation, Sephadex G-150 gel filtration and carboxymethyl-sepharose ion exchange chromatography with a 13-fold increase in specific activity. The apparent molecular weight of P.dig-protease was estimated to be 120 kDa by native high performance liquid chromatography (HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed a single polypeptide at about 30 kDa that indicates a tetrameric protein. The proteolytic activity was inhibited by phenylmethylsulfonyl fluoride suggesting a serine-protease enzyme. P.dig-protease stability was investigated over broad range of pH, temperature, salt concentrations, surfactants and metal ions. The purified P.dig-protease was used for the production of bioactive peptides. Red scorpionfish (Scorpaena notata) muscle was hydrolyzed with P.dig-protease in order to obtain peptides with biological activities. Interestingly, the hydrolysate revealed the presence of antioxidant and angiotensin-I converting enzyme inhibitor peptides. |
| Databáze: | OpenAIRE |
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