PEG/NaPA aqueous two-phase systems for the purification of proteases expressed by Penicillium restrictum from Brazilian Savanna |
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| Institution: | 1. Laboratoire d''étude de la chimiosensibilité du paludisme, Fédération des laboratoires, Hôpital Principal de Dakar, Dakar, Senegal;2. Unité de Parasitologie et d''Entomologie, Département des Maladies Infectieuses, Institut de Recherche Biomédicale des Armées, Marseille, France;3. Aix Marseille Université, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, UM 63, CNRS 7278, IRD 198, Inserm 1095, Institut hospitalo-universitaire en infectiologie, Marseille, France;4. Centre national de référence du Paludisme, Marseille, France;5. Direction Interarmées du Service de Santé, Cayenne, Guyane, France;6. Laboratoire de Parasitologie, Institut Pasteur de la Guyane, Cayenne, Guyane, France;7. Service des Urgences, Hôpital Principal de Dakar, Dakar, Senegal;8. Centre Medical Interarmées, Dakar, Senegal;9. Laboratoire de Bactériologie Virologie, Université Cheikh Anta Diop, CHU Le Dantec, Dakar, Senegal;10. Service de Pédiatrie, Hôpital Principal de Dakar, Dakar, Senegal;11. Service de Réanimation Médicale, Hôpital Principal de Dakar, Dakar, Senegal;12. Laboratoire d''Analyses Médicales, Institut Pasteur, Dakar, Senegal;13. Chefferie, Hôpital Principal de Dakar, Dakar, Senegal;1. Department of Chemical Engineering, Beijing Institute of Petro-chemical Technology, Beijing 102617, China;2. National Key Lab of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China |
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| Abstract: | The partitioning of proteases expressed by Penicillium restrictum from Brazilian Savanna in an inexpensive aqueous two-phase system composed of poly (ethylene glycol) (PEG) and sodium polyacrylate (NaPA) was studied. The effects of PEG molecular weight and concentration, as well as NaPA concentration and the concentration of fermented broth on protease partitioning were studied. Partitioning into the top PEG-rich phase was increased in systems with smaller PEG-molecular weight, higher NaPA concentration and lower PEG concentration. For most systems studied, purification has been achieved by directing the biomolecule partition to the opposite phase of the other proteins, providing the enzyme purification. The highest partition coefficient was obtained using 20 wt% NaPA, 4 wt% PEG 2000 g mol−1 and 45 wt% fermented broth, leading to a purification factor of 1.98 and partition coefficient of 37.73. The system showed high mass balances and yield, indicating enzyme stability and applicability for industrial processes. The partitioning results using the PEG/NaPA/NaCl system show that this method could be used to purify or concentrate protease from fermented broth. |
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| Keywords: | Protease Aqueous two-phase systems Poly (ethylene glycol) Poly (acrylic acid) |
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