Characterization of proteolytic enzyme secreted by Streptomyces cinereoruber ssp. cinereoruber isolated from human pleural fluid
Journal Title: International journal of Advanced Biological and Biomedical Research - Year 2016, Vol 5, Issue 1
Abstract
Actinomycetes are an uncommon agent of human infections and its pathogenic factors are not known. The present study reports a rare case isolation of an actinomycete from a woman pleural fluid; the strain was identified by 16S rRNA gene sequence analysis. This strain was tested to produce an extracellular protease that hydrolysis gelatin, casein and hemoglobin on agar mediums. The purification of the enzyme was carried by ammonium sulfate precipitation, gel filtration and ion exchange chromatographies. The activity of protease was studied at different pH values and temperatures and in the presence of metallic ions and inhibitors. The molecular weight of the enzyme was determined by 12% Tricine SDS-polyacrylamide gel electrophoresis. The strain was identified as Streptomyces cinereoruber ssp. cinereoruber. Extracellular proteolytic enzyme was purified at 19.67 fold and a 3.0% recovery. The enzyme was characterized as having optimal activities at pH 11.0 and 50°C, it keeps more than 50% of activity at pH between 4.0 to 12.0 and it is thermostable at 30 and 40°C. Enzymatic activity is enhanced in the presence of metal ions and inhibited by EDTA and 1,10-phenanthroline. The molecular weight was 53 kDa. This study reports the first case isolation of Streptomyces cinereoruber ssp. cinereoruber from pleural fluid, the extracellular zinc-metalloprotease was proposed as candidate virulence factor.
Authors and Affiliations
Habiba Zerizer| Laboratoire de Génie Microbiologique et Applications, Université des frères Mentouri, Constantine, Algérie, Fateh Merouane| Laboratoire de Génie Microbiologique et Applications, Université des frères Mentouri, Constantine, Algérie, Chahrazad Bougherara| Laboratoire Bioqual, Institut de la Nutrition, de l’Alimentation et des Technologies Agro-Alimentaires, Université des frères Mentouri, Constantine, Algérie, Mouhamed Gagaoua| Laboratoire Bioqual, Institut de la Nutrition, de l’Alimentation et des Technologies Agro-Alimentaires, Université des frères Mentouri, Constantine, Algérie, Kahina Hafid| Laboratoire Bioqual, Institut de la Nutrition, de l’Alimentation et des Technologies Agro-Alimentaires, Université des frères Mentouri, Constantine, Algérie, Faiza Boughachiche| Laboratoire de Génie Microbiologique et Applications, Université des frères Mentouri, Constantine, Algérie, Didier Raoult| Unité des Rickettsies, Faculté de Médecine, Université de la Méditerranée, Marseille, France, Abderrahmane Boulahrouf| Laboratoire de Génie Microbiologique et Applications, Université des frères Mentouri, Constantine, Algérie
Keywords
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- EP ID EP13568
- DOI 10.18869/IJABBR.2016.275
- Views 444
- Downloads 33
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