Hen egg white as a feeder protein for lipase immobilization |
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| Institution: | 1. Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA;2. Texas A&M AgriLife Research Center, Weslaco, TX, USA;1. Pathology and Medical Biology, University Medical Center Groningen and University of Groningen, The Netherlands;2. Nephrology, University Medical Center Groningen and University of Groningen, The Netherlands;3. Department of Clinical Research, University of Bern, Inselspital, Switzerland;4. Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, United Kingdom;5. Top Institute Food and Nutrition, Wageningen, The Netherlands;1. Department of Chemistry & Centre BioMed, Université du Québec à Montréal, CP 8888, Branch A, Montreal, Québec H3C 3P8, Canada;2. Department of Biochemical Sciences, “A. Rossi-Fanelli”, Sapienza University of Rome, P.le A. Moro 5, 00185 Rome, Italy;1. Department of Food Science and Technology, Faculty of Technology and Community Development, Thaksin University, Phatthalung Campus, Phatthalung, 93210, Thailand;2. Department of Food Technology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand;1. Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal;2. Instituto de Ciência e Engenharia de Materiais e Superficies (ICEMS), Instituto Superior Técnico, Universidade Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal;3. Institute for Biotechnology and Bioengineering (IBB), Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Universidade Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal;4. Faculdade de Engenharia, Universidade Lusófona de Humanidades e Tecnologias, 1749-024 Lisboa, Portugal |
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| Abstract: | Enzyme stabilization via immobilization is one of the preferred processes as it provides the advantages of recovery and reusability. In this study, Thermomyces lanuginosus lipase has been immobilized through crosslinking using 2% glutaraldehyde and hen egg white, as an approach towards CLEA preparation. The immobilization efficiency and the properties of the immobilized enzyme in terms of stability to pH, temperature, and denaturants was studied and compared with the free enzyme. Immobilization efficiency of 56% was achieved with hen egg white. The immobilized enzyme displayed a shift in optimum pH towards the acidic side with an optimum at pH 4.0 whereas the pH optimum for free enzyme was at pH 6.0. The immobilized enzyme was stable at higher temperature retaining about 83% of its maximum activity as compared to the free enzyme retaining only 41% activity at 70 °C. The denaturation of lipase in free form was rapid with a half-life of 2 h at 60 °C and 58 min at 70 °C as compared to 12 h at 60 °C and 2 h at 70 °C for the immobilized enzyme. The effect of denaturants, urea and guanidine hydrochloride on the free and immobilized enzyme was studied and the immobilized enzyme was found to be more stable towards denaturants retaining 74% activity in 8 M urea and 98% in 6 M GndHCl as compared to 42% and 33% respectively in the case of free enzyme. The apparent Km (2.08 mM) and apparent Vmax (0.95 μmol/min) of immobilized enzyme was lower as compared to free enzyme; Km (8.0 mM) and Vmax (2.857 μmol/min). The immobilized enzyme was reused several times for the hydrolysis of olive oil. |
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