Purification of serratiopeptidase from Serratia marcescens NRRL B 23112 using ultrasound assisted three phase partitioning |
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| Institution: | 1. Equipe MaQuaV, INATAA, Université Frères Mentouri Constantine, Route de Ain El-Bey, 25000 Constantine, Algeria;2. INATAA, Université Frères Mentouri Constantine, Route de Ain El-Bey, 25000 Constantine, Algeria;3. Laboratory of Applied Microbiology, Faculty of Nature Science and Life, University of Bejaia, Targa Ouzemmour, 06000 Bejaia, Algeria;4. Laboratory of Cellular and Molecular Biology, Microbiology Team, University of Sciences and Technology of Houari Boumediene, PO Box 32, El Alia, Bab Ezzouar, 16111 Algiers, Algeria;1. Department of Clinical Microbiology, Royal College of Surgeons in Ireland, Beaumont Hospital, Dublin 9, Ireland;2. Department of Microbiology, Beaumont Hospital, Dublin 9, Ireland;3. Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland;4. Department of Microbiology, Connolly Hospital, Dublin 15, Ireland;1. Teagasc Food Research Centre, Dublin, Ireland;2. Food Packaging Group, School of Food and Nutritional Sciences, University College Cork, Cork, Ireland |
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| Abstract: | The ultrasound assisted three phase partitioning (UATPP) is a novel bioseparation method for separation and purification of biomolecules. In the present work, UATPP was investigated for the first time for purification of serratiopeptidase from Serratia marcescens NRRL B 23112. Effect of various process parameters such as ammonium sulphate saturation, t-butanol to crude extract ratio, pH, ultrasonic frequency, ultrasonic intensity, duty cycle and irradiation time were evaluated and optimized. The optimized conditions were found to be as follows: ammonium sulphate saturation 30% (w/v), pH 7.0, t-butanol to crude ratio 1:1 (v/v), ultrasound frequency 25 kHz, ultrasound intensity 0.05 W/cm2, duty cycle 20% and irradiation time 5 min. The maximum purity and recovery obtained from UATPP was 9.4-fold and 96% respectively as compared to the three phase partitioning (TPP) (4.2-fold and 83%). Also the process time for UATPP was significantly reduced to 5 min from 1 h as compared to TPP. The results indicate that, UATPP is an efficient technique for the purification of serratiopeptidase with maximum purity, recovery and reduced processing time. |
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| Keywords: | Ultrasound assisted three phase partitioning (UATPP) Serratiopeptidase Three phase partitioning (TPP) Purification |
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