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Operational and Thermal Stability Analysis of <Emphasis Type="Italic">Thermomyces lanuginosus</Emphasis> Lipase Covalently Immobilized onto Modified Chitosan Supports
Authors:Horacio L Bonazza  Ricardo M Manzo  José C S dos Santos  Enrique J Mammarella
Institution:1.Instituto de Desarrollo Tecnológico para la Industria Química (INTEC), Universidad Nacional del Litoral (UNL),Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET),Santa Fe,Argentina;2.Instituto de Engenharias e Desenvolvimento Sustentável,Universidade da Integra??o Internacional da Lusofonia Afro-Brasileira,Acarape,Brazil
Abstract:The aim of this paper was to evaluate different strategies of chitosan activation using cross-linking reagent like glycidol, epichlorohydrin, and glutaraldehyde for Thermomyces lanuginosus lipase (TLL) immobilization. Operational activity and stability by esterification of oleic acid with ethanol and thermal inactivation using these derivatives were investigated. Derivative obtained by sequentially activation with glycidol, ethylenediamine, and glutaraldehyde and subsequent TLL immobilization showed the best performance, with high hydrolytic activity value. Its stability was 15-fold higher than solubilized TLL in the evaluated inactivation conditions (60 °C, 25 mM sodium phosphate buffer pH 7). After 5 cycles of oleic acid esterification, only a few percentage of its conversion has reduced. On the other hand, glycidol-activated chitosan derivative showed very low hydrolytic activity value. Epichlorohydrin-activated chitosan derivative showed regular hydrolytic activity value. Both derivatives showed low immobilization yields. Operational stability of this last derivative was very low, where after the first cycle of oleic acid esterification, only 56% of its initial conversion was obtained.
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