One-step separation of lysozyme by reverse micelles formed by the cationic surfactant,cetyldimethylammonium bromide |
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| Affiliation: | 1. Instituto de Ciência e Tecnologia, UFVJM, Rodovia MGT 367, Km 583, n° 5000, Alto do Jacuba, 391000-000 Diamantina, MG, Brazil;2. Departamento de Química, UFVJM, Rodovia MGT 367, Km 583, n° 5000, Alto do Jacuba, 391000-000 Diamantina, MG, Brazil;3. Instituto de Química, UNICAMP, P.O. Box 6154, 13084-971 Campinas, SP, Brazil;4. Instituto Nacional de Ciência e Tecnologia em Bioanalítica, Unicamp, 13083-970 Campinas, SP, Brazil;1. Gause Institute of New Antibiotics, Russian Academy of Medical Sciences, 11 B. Pirogovskaya Street, Moscow 119021, Russia;2. Mendeleyev University of Chemical Technology, 9 Miusskaya Square, Moscow 125190, Russia;3. Blokhin Cancer Center, 24 Kashirskoye Shosse, Moscow 115478, Russia;4. Moscow Engineering and Physics Institute, 31 Kashirskoye Shosse, Moscow 115409, Russia;5. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov Street, Moscow 119991, Russia;6. Rega Institute for Medical Research, KU Leuven, 3000 Leuven, Belgium;7. Developmental Therapeutics Branch, National Cancer Institute, NIH, 37 Convent Drive, 37-5068, Bethesda, MD 20892, USA;1. Key Laboratory of UV-Emitting Materials and Technology (Northeast Normal University), Ministry of Education, Changchun, Jilin, 130024, China;2. School of Life Sciences, Northeast Normal University, Changchun, Jilin, 130024, China;1. Energy Materials & Surface Science Laboratory, Solar Energy Research Center, School of Chemical Engineering, Chonbuk National University, Jeonju 54896, Republic of Korea;2. New & Renewable Energy Material Development Center (NewREC), Chonbuk National University, Jeonbuk, Republic of Korea;1. Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, United States;2. Department of Physics, University of Michigan, Ann Arbor, MI 48109, United States |
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| Abstract: | Lysozyme was selectively extracted from reconstituted freeze-dried egg-white, using reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide (CDAB). The major egg-white proteins, including ovalbumin and ovotansferrin, were solubilized into the organic phase while lysozyme was recovered in the aqueous phase. The solubilization behaviours of proteins were manipulated by processing parameters, including pH and salt concentration in the aqueous phase and concentration of surfactant in the organic phase. The optimum extraction was achieved with sodium borate buffer (50 mM, pH 9, no added KCl) and organic phase containing 50 mM CDAB. After the forward extraction, 96% of total lysozyme activity was recovered. Lysozyme was efficiently purified, more than 30-fold with only a single forward extraction. The suggested extraction procedure has advantages in terms of time and cost compared to traditional reverse micellar extraction which requires both forward and backward extraction steps. |
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