Optimization of tetramethoxysilane-derived sol gel entrapment protocol stabilizes highly active chlorophyllase |
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Authors: | Yunyu Yi Ronald Neufeld Selim Kermasha |
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Institution: | (1) Department of Chemical Engineering, Queen’s University, Kingston, Ont., Canada, K7L 3N6;(2) Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste-Anne de Bellevue, Quebec, Canada, H9X 3V9 |
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Abstract: | Entrapment of membrane proteins is a challenging task compared to that involving soluble proteins. Chlorophyllase, a membrane
protein, was successfully entrapped in tetramethoxysilane-derived sol-gel. Pre-gel sol typically consists of an aqueous suspension
of chlorophyllase, precursors including tetramethoxysilane and/or methytrimethoxysilane, and sodium fluoride as catalyst.
To obtain a highly active entrapped enzyme preparation, the effects of various immobilization parameters, including the chemical
compositions of pre-gel sol (water/silane ratio, precursor type and proportions, enzyme loading, sodium fluoride concentration),
and sol-gel process parameters (aging and drying time and approach) have been investigated. Chlorophyllase demonstrated the
highest activity in gel derived from a pre-gel sol with water/silane ratio of 30 and enzyme loading of 0.257 mgprotein/ggel, and showed moderately lower activity in organically modified sol-gel than that in hydrophilic sol-gel. The effects of water/silane
ratio and precursor combinations on the activity of entrapped chlorophyllase were also studied by examining the pore morphology
of gel via nitrogen adsorption-desorption. Longer aging time leads to an entrapped chlorophyllase preparation with higher
activity. Chlorophyllase preparation demonstrated negligible activity after air-drying for 12 h while lyophilized chlorophyllase
preparation demonstrated 8, 4 and 4 times higher activity than air-dried, vacuum-dried and solvent-dried preparations. Chlorophyllase
demonstrated 30% higher activity in the improved sol-gel protocol than that from a non-optimized sol-gel protocol developed
in a previous study. |
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Keywords: | Chlorophyllase Membrane protein Encapsulation Sol-gel chemical composition Sol-gel process parameters |
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