Immobilization of Emphasis Type="Italic"Candida

Authors: Viviane Chiaradia Nicole Spillere Soares Alexsandra Valério Débora de Oliveira Pedro H H Araújo Claudia Sayer

Publish Date: 2016/05/16

Volume: 180, Issue: 3, Pages: 558-575

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Abstract

Polyureaurethane PUU nanoparticles with encapsulated superparamagnetic magnetite Fe3O4 were obtained by interfacial miniemulsion polymerization and used as support for immobilization of lipase B from Candida antarctica CALB CALB enzyme was immobilized on magnetic PUU nanoparticles in two steps The enzyme was immobilized in the lyophilized nanoparticles magnetic PUU after the support synthesis in phosphate buffer pH 76 containing CALB by the contact between nanoparticles and enzymatic solution The mixture was incubated at 30 °C in an orbital shaker during 05 until 6 h to determine the time for maximum immobilization efficiency The enzyme activity was determined by esterification reactions between lauric acid and propanol Residual activities above 95  in relation to free enzyme were obtained in 1 h of immobilization with enzyme concentration of 055 mg/mL FTIR spectrum and SEMFEG images were used to confirm the presence of CALB on magnetic support after immobilization and stability of support even after immobilization process respectively Thermal 40 60 and 80 °C and pH pH 4 7 and 10 stabilities storage stability and reuse were evaluated CALB immobilized derivatives showed high stabilities with residual activities of 95 100 and 100  at 40 60 and 80 °C respectively in 6 h of incubation After incubation in different pH values CALB immobilized derivative presented activities of 81 76 and 69  in relation to activities in the beginning of the stabilization process in pH 4 7 and 10 respectively Furthermore CALB immobilized derivative reduces only 15  of its activity after 30 days of storage at 4 °C Reuse results showed that immobilized CALB on magnetic PUU nanoparticles led to 95  of geranyl oleate conversion after 8 cycles of application demonstrating high stability of the CALB immobilized derivative under different conditionsThe authors would like to thank Conselho Nacional de Desenvolvimento Científico e Tecnológico CNPq and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior CAPES for the financial support and Laboratório Central de Microscopia Eletrônica da UFSC LCMEUFSC for the TEM SEMFEG and fluorescence microscopy analyses

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