Celulase e álcool desidrogenase imobilizadas em filmes de Langmuir -Blodgett para identificação de celulose e etanol no nível molecular

• Autor(a) principal: Rodrigues, Dilmer [UNIFESP]
• Data de Publicação: 2013
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=152359; https://repositorio.unifesp.br/handle/11600/47892
• topic: Sustainability; Energy; Enzymes; Thin films; Langmuir monolayers; Langmuir-Blodgett films; Sustentabilidade; Energia; Enzimas; Filmes finos; Monocamadas de Langmuir; Filmes de Langmuir-Blodgett

In this study, we investigated at the molecular level the incorporation of cellulase and alcohol dehydrogenase (ADH) in Langmuir and Langmuir-Blodgett (LB) films of dipalmitoylphosphatidylcholine (DPPC). The mixed DPPC-enzyme films were then investigated upon contact with cellulose and ethanol. The changes at the molecular level in the properties of the films were analyzed by optical and tensiometric techniques. We aim to obtain information on the production of ethanol in a more efficient manner. The first evidence that enzymes adsorbed at the films in the lipid-water interface was through adsorption kinetic curves, wherein upon insertion of the enzyme in the aqueous subphase for monolayers of DPPC, an increase in the surface pressure values for the phospholipid matrix was observed. The second evidence was the analysis of the surface pressure-area isotherms, which showed that the presence of the enzymes caused the expansion of the lipid monolayer to higher areas. The third evidence was observed with polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS), studied in Langmuir films, which revealed bands in the regions between 1500 cm-1 and 1800 cm-1, related to protein groups, namely amide II ( CN and NH bendings) and amide I (C = O stretch). In the sequence, the films were transferred to solid matrices as LB films. The PM-IRRAS analysis confirmed the successful transfer of enzyme and phospholipids. The molecular interaction between the mixed LB film (DDPC / ADH / cellulase) and cellulose was observed through PM-IRRAS since in the regions between 1500 cm-1 and 3500 cm-1, changes were observed revealing alterations in the secondary structure of the protein, related to beta-sheet, alpha-helix, beta-turn, and unordered structures. Also for the regions between 2800 cm-1 and 3500 cm-1, the symmetric and asymmetric stretches for CH in CH2 CH3 groups, as well as the stretching of amines (NH2) and OH groups could be investigated. All these factors indicate the sensitivity of the enzymes when placed in contact with cellulose dissolved in ionic liquid.