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Perfil proteolítico de extratos aquosos de folha e semente de Mucuna pruriens (L.) DC

Raquel Elisa Silva-López, Nathalia N. M. Vidal, Nathalia M. N. Vidal, Rayane N. Gonçalves, Rayane N. Gonçalves

DOI: http://dx.doi.org/10.5935/2446-4775.20160035

Resumo


Mucuna pruriens (L.) DC é uma planta medicinal, da família Fabaceae, usada no tratamento de diversas doenças. Extratos aquosos de folhas e sementes de M. pruriens foram preparados utilizando água, detergente Triton X-100 e tampões fosfato de sódio e Tris-HCl. Os extratos de folha exibiram os maiores teores de proteínas. Todos os extratos exibiram atividade sobre o substrato N-α-p-tosil-L-arginil-metil éster (L-TAME), principalmente nas faixas de pH ácido (4,0-5,0) e alcalino (9,0- 9,5), e atividade máxima nas temperaturas entre 40-60ºC, com exceção do extrato de semente obtido com tampão Tris-HCl (MP-ST), que apresentou pico em 80ºC. Os extratos de folhas exibiram perfis eletroforéticos semelhantes sob condições redutoras e não redutoras, sendo observada uma proteína majoritária com de cerca de 30 kDa. Na análise capacidade proteolítica, o zimograma apresentou um perfil distinto em diferentes valores de pH: em pH 9,0 os extratos obtidos com água (MP-EA) e com detergente (MP-ED) apresentaram atividade na região próxima de 170 kDa; em pH 9,5 pôde-se observar duas regiões de proteólise entre 66-90 e 200-250 kDa, com exceção do MP-EA. Os extratos de semente apresentaram perfis distintos em condições redutoras e não redutoras, com proteínas majoritárias entre 35 e 23 kDa, e atividade lítica sobre a gelatina na região de 80 kDa. Todos os extratos apresentaram atividade proteolítica contra os substratos proteicos hemoglobina, caseína e albumina e boa estabilidade na presença de agentes surfactantes e oxidantes. As proteases presentes nos extratos de M. pruriens sugerem ser da classe das serino e metaloproteases, com atividade modulada positivamente na presença íons Mn2+ e Ca2+. Essas características bioquímicas conferem aos extratos de M. Pruriens grande valor biotecnológico e possível potencial terapêutico.


Palavras-chave


Mucuna pruriens. Proteases. Atividade enzimática. Estabilidade.

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