Imobilização multipontual das peroxidases da casca da soja e chuchu em suportes alternativos de pó de sabugo de milho e celulose bacteriana
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Data
2012-01-11
Autores
Vinueza Galárraga, Julio César [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
As peroxidases (EC 1.11.1.7) são hemoproteínas que catalisam processos redox e durante a oxidação são gerados radicais livres, formando produtos poliaromáticos insolúveis em água, facilitando sua remoção do meio aquoso. O objetivo deste trabalho foi extrair as peroxidases da casca de soja e do chuchu, realizar aminação na superfície da estrutura terciária das enzimas e imobilizarem estas enzimas nos suportes alternativos de celulose vegetal (pó de sabugo de milho) e de celulose bacteriana ativada e utilizar os derivados obtidos para descoloração do azul de bromofenol 0.01 mM. A quantidade de proteínas nos extratos foram determinada por Bradford com valores médios de 0,235 mg mL -1 para soja e 0,290 mg mL -1 para o chuchu, a atividade específica de peroxidase foi determinada com ABTS 1 mM em presença de H2O2 100 mM em λ= 430nm, obtendo-se os valores médios de 86,06 μmol min -1 mg -1 e 9,04 μmol min -1 mg -1 respectivamente, em tampão acetato de sódio 100 mM, pH 4,0. Aminação das peroxidases solúveis foram feitas em tampão etilenodiamina pH 4,75 e carbodiimida, 10 e 50 mM, a funcionalização das celuloses e imobilização covalente multipontual foram semelhantes às descritas para agarose (Guisan). 19 As peroxidases aminadas 10 e 50 mM foram imobilizadas covalentemente nos suportes ativados SM-glioxil e CB-glioxil. Para a descoloração do azul de bromofenol 0.01 mM, utilizou-se os derivados estabilizados em presença de H2O2 em λ= 590nm. Os derivados foram reutilizados por cinco reciclos, mantendo-se a propriedade catalítica, sugerindo que estes derivados são uma alternativa de baixo custo, não só para o tratamento de águas residuais como também com potenciais usos em outros processos industriais
The peroxidases (EC 1.11.1.7) are hemoproteins that catalyze redox processes; during the oxidation the generated free radicals are forming poly-aromatic products insoluble in water, facilitating their removal from aqueous medium. The objective of this study was to extract peroxidases from the coat soybean and the chuchu (Sechium edule L), to make aminations on the tertiary structure of the enzymes surface, and immobilize these enzymes in the alternative supports of plant cellulose (powdered corn cob) and active bacterial cellulose, to use the derivatives obtained for the discoloration of the bromophenol blue 0.01 mM. The amount of protein in the extracts were determined by Bradford methods, with average values of 0,235 mg mL -1 for soybean and 0,290 mg mL -1 for chuchu peroxidases, the specific activity of peroxidase was determined with ABTS 1 mM in the presence of H2O2 100 mM in λ= 430nm, obtaining the mean values of 86,06 μmol min -1 mg -1 and 9,04 μmol min -1 mg -1 respectively, in sodium acetate buffer 100mM, pH 4,0. Amination of soluble peroxidases were made in buffer ethylenediamine pH 4,75 and carbodiimide, 10 and 50mM, the functionalization of cellulose and multipoint covalent immobilization were similar to those described for agarose (Guisan). 19 The amino peroxidases 10 and 50 mM were covalently immobilized on activated supports SM-glioxil and CB-glioxil. For discoloration of bromophenol blue 0.01 mM, we used derivatives of stabilized in H2O2 presence in λ= 590nm. The derivatives were reused for five recycling maintaining the catalytic property, suggesting that these products are a low cost alternative, not only for the treatment of wastewater as well as potential uses in other industrial processes
The peroxidases (EC 1.11.1.7) are hemoproteins that catalyze redox processes; during the oxidation the generated free radicals are forming poly-aromatic products insoluble in water, facilitating their removal from aqueous medium. The objective of this study was to extract peroxidases from the coat soybean and the chuchu (Sechium edule L), to make aminations on the tertiary structure of the enzymes surface, and immobilize these enzymes in the alternative supports of plant cellulose (powdered corn cob) and active bacterial cellulose, to use the derivatives obtained for the discoloration of the bromophenol blue 0.01 mM. The amount of protein in the extracts were determined by Bradford methods, with average values of 0,235 mg mL -1 for soybean and 0,290 mg mL -1 for chuchu peroxidases, the specific activity of peroxidase was determined with ABTS 1 mM in the presence of H2O2 100 mM in λ= 430nm, obtaining the mean values of 86,06 μmol min -1 mg -1 and 9,04 μmol min -1 mg -1 respectively, in sodium acetate buffer 100mM, pH 4,0. Amination of soluble peroxidases were made in buffer ethylenediamine pH 4,75 and carbodiimide, 10 and 50mM, the functionalization of cellulose and multipoint covalent immobilization were similar to those described for agarose (Guisan). 19 The amino peroxidases 10 and 50 mM were covalently immobilized on activated supports SM-glioxil and CB-glioxil. For discoloration of bromophenol blue 0.01 mM, we used derivatives of stabilized in H2O2 presence in λ= 590nm. The derivatives were reused for five recycling maintaining the catalytic property, suggesting that these products are a low cost alternative, not only for the treatment of wastewater as well as potential uses in other industrial processes
Descrição
Palavras-chave
Soja - Pesquisa, Milho - Pesquisa, Chuchu - Pesquisa, Peroxidases - Coat soybean
Como citar
VINUEZA GALÁRRAGA, Julio César. Imobilização multipontual das peroxidases da casca da soja e chuchu em suportes alternativos de pó de sabugo de milho e celulose bacteriana. 2012. 103 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2012.