Extração e purificação da bromelina a partir do abacaxi para a hidrólise do glúten
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Citações na Scopus
Tipo de produção
Trabalho de Conclusão de Curso
Data
2023-06-15
Autores
Andrade, Ana Carolina
Vendramini, Beatriz
Neves, Julia Toledo
Martinelli, Sabrina
Vendramini, Beatriz
Neves, Julia Toledo
Martinelli, Sabrina
Orientador
Lucarini, Adriana Célia
Periódico
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Texto completo (DOI)
Palavras-chave
bromelina,abacaxi,extração líquido-líquido,bromelin,pineapple,liquid-liquid extraction
Resumo
A Bromelina é uma enzima proteolítica de origem vegetal obtida em diversas espécies da família Bromeliaceae, sendo principalmente encontrada no abacaxi. Ela é utilizada em diversos setores, dentre eles se destacam o tratamento de edemas, na digestão de proteínas e no amaciamento de carnes. Neste trabalho, foi realizado um estudo visando a purificação da bromelina do fruto do abacaxi. A purificação da bromelina foi realizada por sistema de extração líquido-líquido em duas fases aquosas 20 % (m/m) de PEG e 15 % (m/m) de fosfato. Diferentes massas molares de PEG (400, 600, 1500 e 4000) e valores de pH (6,0 / 7,0 / 8,0) foram testados. Os resultados obtidos mostraram que as extrações com PEG 400, 600 e 1500 não levaram a uma efetiva separação e purificação das proteínas, resultando em um aumento de pureza próximo de 1,0; exceção para PEG 1500 pH 8 que mostrou aumento de pureza e recuperação da bromelina na fase inferior de 100%. Em todos estes estudos a bromelina migrou preferencialmente para a fase inferior (fosfato). No entanto, a extração com PEG 4000 mostrou-se mais eficiente, com uma eficaz separação das fases nos três valores de pH e uma recuperação de atividade enzimática praticamente total na fase inferior. Os valores de aumento de pureza evidenciaram a eficácia da purificação nos valores de pH 6,0 e 8,0, com um aumento de pureza de cerca de duas vezes. Além disso, estudou-se a etapa de precipitação com sulfato de amônio a 60 e 80% de saturação para clarificação do extrato de abacaxi. Os estudos mostraram que o precipitado obtido a 80% de saturação e solubilizado apresentou uma recuperação de atividade próximo de 77%, confirmando a viabilidade dessa etapa para aumentar a concentração e promover a clarificação do extrato, pois não houve aumento de pureza. Efetuando-se um processo conjugado com duas etapas em série de precipitação do extrato com 80% de saturação de sulfato de amônio, seguida de extração líquido-líquido em sistema de duas fases aquosas utilizando PEG 4000 e fosfato em pH 6,0 obteve-se um aumento de pureza na amostra em cerca de 4 vezes e uma recuperação da atividade enzimática em 100% na fase inferior. Os dados obtidos neste estudo fornecem informações relevantes para o desenvolvimento de processos de purificação da bromelina a partir do abacaxi, contribuindo para possibilitar sua utilização em aplicações terapêuticas e industriais.
Bromelin is a proteolytic enzyme of plant origin obtained from several species of the Bromeliaceae family, mainly found in pineapple. It is used in several areas, among them are the treatment of edema, protein digestion, and meat tenderization. In this work, a study was conducted to purify bromelain from the pineapple fruit. The purification of bromelain was performed by liquid-liquid extraction system in two aqueous phases 20 % (w/w) PEG and 15 % (w/w) phosphate. Different PEG molar masses (400, 600, 1500 and 4000) and pH values (6.0 / 7.0 / 8.0) were tested. The results obtained showed that the extractions with PEG 400, 600 and 1500 did not lead to an effective separation and purification of the proteins, resulting in an increase in purity close to 1.0; except for PEG 1500 pH 8 which showed an increase in purity and recovery of bromelain in the lower phase of 100%. In all these studies bromelin migrated preferentially to the lower (phosphate) phase. However, the extraction with PEG 4000 proved to be more efficient, with an effective separation of the phases in all three pH values and a practically total recovery of enzyme activity in the lower phase. The values of purity increase evidenced the effectiveness of purification at pH values 6.0 and 8.0, with an increase in purity of about twofold. In addition, the precipitation step with ammonium sulfate at 60 and 80% saturation was studied to clarify the pineapple extract. The studies showed that the precipitate obtained at 80% saturation and solubilized showed a recovery of activity close to 77%, confirming the feasibility of this step to increase the concentration and promote clarification of the extract, because there was no increase in purity. In a conjugated process with two steps in series of precipitation of the extract with 80% saturation of ammonium sulfate, followed by liquid-liquid extraction in a two-phase aqueous system using PEG 4000 and phosphate at pH 6.0, an increase in sample purity of about 4 times and a recovery of enzyme activity of 100% in the lower phase was obtained. The data obtained in this study provide relevant information for the development of purification processes of bromelain from pineapple, helping to enable its use in therapeutic and industrial applications.
Bromelin is a proteolytic enzyme of plant origin obtained from several species of the Bromeliaceae family, mainly found in pineapple. It is used in several areas, among them are the treatment of edema, protein digestion, and meat tenderization. In this work, a study was conducted to purify bromelain from the pineapple fruit. The purification of bromelain was performed by liquid-liquid extraction system in two aqueous phases 20 % (w/w) PEG and 15 % (w/w) phosphate. Different PEG molar masses (400, 600, 1500 and 4000) and pH values (6.0 / 7.0 / 8.0) were tested. The results obtained showed that the extractions with PEG 400, 600 and 1500 did not lead to an effective separation and purification of the proteins, resulting in an increase in purity close to 1.0; except for PEG 1500 pH 8 which showed an increase in purity and recovery of bromelain in the lower phase of 100%. In all these studies bromelin migrated preferentially to the lower (phosphate) phase. However, the extraction with PEG 4000 proved to be more efficient, with an effective separation of the phases in all three pH values and a practically total recovery of enzyme activity in the lower phase. The values of purity increase evidenced the effectiveness of purification at pH values 6.0 and 8.0, with an increase in purity of about twofold. In addition, the precipitation step with ammonium sulfate at 60 and 80% saturation was studied to clarify the pineapple extract. The studies showed that the precipitate obtained at 80% saturation and solubilized showed a recovery of activity close to 77%, confirming the feasibility of this step to increase the concentration and promote clarification of the extract, because there was no increase in purity. In a conjugated process with two steps in series of precipitation of the extract with 80% saturation of ammonium sulfate, followed by liquid-liquid extraction in a two-phase aqueous system using PEG 4000 and phosphate at pH 6.0, an increase in sample purity of about 4 times and a recovery of enzyme activity of 100% in the lower phase was obtained. The data obtained in this study provide relevant information for the development of purification processes of bromelain from pineapple, helping to enable its use in therapeutic and industrial applications.