Effect of high salt concentrations on the stability of immobilized lipases: Dramatic deleterious effects of phosphate anions |
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| Institution: | 1. Departamento de Biocatálisis, ICP-CSIC, Campus UAM-CSIC, Madrid, Spain;2. Food Biotechnology Division, Biotechnology Research Center (CRBt), Constantine, Algeria;3. Faculty of Nature and life Sciences, Ibn Khalboun University, Tiaret, Algeria;4. Heterogenous Biocatalysis laboratory, CIC Biomagune, Paseo Miramon 182, 20014, San Sebastian, Spain;5. IKERBASQUE, Basque Foundation for Science, Bilbao, Spain;1. Department of Bioengineering and IBB—Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal;2. Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal;3. Centro de Química-Física Molecular and IN, Instituto Superior Técnico, Universidade de Lisboa, Portugal;4. Faculty of Engineering, Universidade Lusófona, Lisboa, Portugal;1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30, Puzhu South Road, Nanjing, 211816, China;2. National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, China;3. National Center for Protein Science Shanghai, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 333 Haike Road, Shanghai, 201210, China;4. Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China;5. Jiangsu National Synergetic Innovation Center for Advanced Materials, Nanjing, 211816, China;6. Jiangsu Industrial Technology Research Institute, Nanjing, China;1. Grupo de Biocatálisis, Planta Piloto de Ingeniería Química (PLAPIQUI), Universidad Nacional del Sur – CONICET, Camino La Carrindanga Km 7 CC 717, 8000 Bahía Blanca, Argentina;2. Instituto de Tecnología en Polímeros y Nanotecnología (ITPN), Facultad de Ingeniería, Universidad de Buenos Aires – CONICET, Las Heras 2214, 1127AAR Buenos Aires, Argentina;1. Department of Chemistry, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India;2. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India;1. Soft Matter Nanotechnology Group, CIC biomaGUNE, Paseo Miramón 182C, 20009 San Sebastián, Guipúzcoa, Spain;2. Instituto de Investigaciones Fisicoquimicas Teóricas y Aplicadas (INIFTA), (UNLP, CONICET), Sucursal 4, Casilla de Correo 16, 1900 La Plata, Argentina;3. Experimental Physics, Saarland University, 66123 Sarbruecken, Germany;1. Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain;2. Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale, Universita’ degli Studi di Napoli Federico II, Italy;3. Unidad de Investigación y Desarrollo en Alimentos, Instituto Tecnológico de Veracruz, Calzada Miguel A. de Quevedo 2779, 91897 Veracruz, Mexico;4. Istituto di Ricerche sulla Combustione—Consiglio Nazionale delle Ricerche, Napoli, Italy |
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| Abstract: | We have analyzed the effects of the buffer nature on the stability of immobilized lipases. Commercial phospholipase Lecitase Ultra (LU), lipase B from Candida antarctica (CALB) and lipase from Thermomyces lanuginosus (TLL) have been immobilized on octyl-glyoxyl agarose beads. The enzymes were readily inactivated using 4 M sodium phosphate but 6 M NaCl did not inactivate them. Using 2 M of sodium phosphate, the inactivation of the 3 immobilized enzymes still was very significant even at 25 °C but at lower rate than with higher phosphate concentration. Thermal stress inactivations of the immobilized enzymes revealed that even 100 mM sodium phosphate produced a significant decrease in enzyme stability; this effect was less pronounced for Lecitase but dramatic for CALB. While 6 M NaCl presented slightly positive (LU) or negative (TLL) effects on their thermal stabilities of, CALB was thermally stabilized under the same conditions. Results were very different using free enymes. Fluorescence spectroscopy revealed dramatic structural rearrangements of the immobilized enzymes in the presence of high phosphate concentration. From these results, the use of sodium phosphate does not seem to be recommended for studies on thermal stability of lipases, although this should be verified for each enzyme and immobilized preparation. |
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| Keywords: | Lipase stability Phosphate inactivation of lipases Buffers for lipases Ionic strength and enzyme stability Lipases immobilized in heterofunctional supports |
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