Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279
DOI:
https://doi.org/10.20873/jbb.uft.cemaf.v3n1.faroukPalavras-chave:
Enterobacter sakazakii, enzima degradadora de fitato, fitato, purificaçãoResumo
Uma enzima extracelular degradadora de fitato produzida por Enterobacter sakazakii ASUIA279 foi purificada para homogeneidade usando cromatografia de troca aniônica por FPLC e filtração em gel. A enzima foi purificada cerca de 66 vezes com uma recuperação de 27%. Sua massa molecular foi estimada em 43 kDa por SDS-PAGE. A constante de Michaelis (KM) e o número de rotatividade (kcat) para o fitato de sódio a pH 5,0 e 50 ° C foram calculados a partir do gráfico de Lineweaver-Burk como 760 µM e 4,14s-1, respectivamente. A enzima mostrou especificidade estreita do substrato e não fitato, mas o GTP foi desfosforilado com a maior taxa relativa de hidrólise. No entanto, de acordo com os valores de kcat / KM, concluiu-se que o fitato é o substrato in vivo da enzima. A atividade ideal foi determinada em pH 4,5 e 45-55 ° C. A enzima foi fortemente inibida por Fe3 +, Cu2 +, Zn2 +, molibdato, vanadato, fluoreto e fosfato (1 mM).
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