Electrostatic and Water Content Effects on Yarrowia lipolytica Lipase Immobilization by Adsorption
Wazé A. Mireille Alloue-Boraud *
Walloon Centre of Industrial Biology (CWBI) Unit of Bio-Industries, University of Liege, Gembloux Agro-Bio Tech, Passage des Deportees 2, B-5030 Gembloux, Belgium and Laboratory of Microbiology and Molecular Biology, UFR-STA, University of Nangui Abrogoua former, University of Abobo Adjamé, 02 BP 801 Abidjan 02, Côte d’Ivoire
Annick Lejeune
Walloon Centre of Industrial Biology (CWBI) Unit of Bio-Industries, University of Liege, Gembloux Agro-Bio Tech, Passage des Deportees 2, B-5030 Gembloux, Belgium
Rose Koffi-Nevry
Laboratory of Microbiology and Molecular Biology, UFR-STA, University of Nangui Abrogoua former, University of Abobo Adjamé, 02 BP 801 Abidjan 02, Côte d’Ivoire
Jacqueline Destain
Walloon Centre of Industrial Biology (CWBI) Unit of Bio-Industries, University of Liege, Gembloux Agro-Bio Tech, Passage des Deportees 2, B-5030 Gembloux, Belgium
Michel Paquot
Unit of Biological Chemistry and Industrial, University of Liege, Gembloux Agrobio-Tech, Passage des Deportees 2, B-5030 Gembloux, Belgium
Philippe Thonart
Walloon Centre of Industrial Biology (CWBI) Unit of Bio-Industries, University of Liege, Gembloux Agro-Bio Tech, Passage des Deportees 2, B-5030 Gembloux, Belgium
*Author to whom correspondence should be addressed.
Abstract
Aims: In the present investigation, an attempt has been made to explain lipase immobilization by adsorption on three minerals matrixes, i.e. Celite 545, Silica gel (60G) and Avicel (PH 101).
Study Design: immobilization by absorption on minerals matrixes, water content by volumetric karl Fischer titration and surface potentials using a particle charge detector Mutek PCD 03 were used.
Place and Duration of Study: Walloon Centre of Industrial Biology (CWBI) Unit of Bio-Industries, University of Liege, Gembloux Agro-Bio Tech, Passage des Deportes 2, B-5030 Gembloux, Belgium between Jun 2012 and jun 2013.
Methodology: A methodical order was developed whereby the influences of water content, surface potentials and pH, on immobilization by adsorption were explored. Adsorbed YLL was used to understand an interesterification reaction between rapeseed oil and milk fat in comparison with a commercial silica-granulated Thermomyces lanuginosus lipase (Lipozyme TL IM).
Results: Maximum immobilization yield was obtained with Celite (70%) and the lowest with silica gel (29%). Total water content of free and immobilized lipase was determined by volumetric Karl Fischer titration. The water content of Silica gel was higher than the one of other supports. Water content of silica gel could prevent the enzyme fixation. These results could be explained by the adsorption being governed mainly by electrostatic interactions between the enzyme and matrix. This hypothesis was further reinforced by measurements of electrical potential. They showed a lowest negative potential of Silica gel after enzyme adsorption in comparison to Celite.
Conclusion: From these results celite was designated as an efficient matrix to immobilize Yarrowia lipolytica lipase (YLL) by adsorption. This performed system was used to realize an interesterification reaction between rapeseed oil and milk fat in comparison with a commercial silica-granulated Thermomyces lanuginosus lipase (Lipozyme TL IM).
Keywords: Yarrowia lipolytica lipase, celite, silica gel, avicel, immobilization, water content, electrostatic interaction, interesterification