Purificação e propriedades de uma enzima degradadora de fitato produzida por Enterobacter sakazakii ASUIA279

Authors

  • Universidade Federal do Tocantins
  • Abd El Aziem Farouk Faculty of Science - Taif University
  • Ralf Greiner Max Rubner Institut - Haid und Neu Strasse, Germany
  • Anis Shobirin Meor Hussin University Putra Malasya

DOI:

https://doi.org/10.20873/jbb.uft.cemaf.v3n1.farouk

Keywords:

Enerobacter sakazakii, phytate-degrading enzyme, phytate, purification

Abstract

An extracellular phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279 was purified to homogeneity using FPLC anion exchange chromatography and gel filtration. The enzyme was purified about 66-fold with a recovery of 27%. Its molecular mass was estimated to be 43 kDa by SDS-PAGE. The Michaelis constant (KM) and turnover number (kcat ) for sodium phytate at pH 5.0 and 50°C were calculated from the Lineweaver-Burk plot to be 760 µM and 4.14s-1, respectively. The enzyme showed narrow substrate specificity and not phytate, but GTP was dephosphorylated with the highest relative rate of hydrolysis. However, according to the kcat/KM values, phytate was concluded to be the in vivo substrate of the enzyme. Optimal activity was determined at pH 4.5 and 45-55°C. The enzyme was strongly inhibited by Fe3+, Cu2+, Zn2+, molybdate, vanadate, fluoride and phosphate (1 mM).

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Published

2012-02-15

How to Cite

Abd-ElAziem, Ralf, Anis Shobirin Meor, Farouk, A. E. A., Greiner, R., & Hussin, A. S. M. (2012). Purificação e propriedades de uma enzima degradadora de fitato produzida por Enterobacter sakazakii ASUIA279. Journal of Biotechnology and Biodiversity, 3(1), 1–9. https://doi.org/10.20873/jbb.uft.cemaf.v3n1.farouk