Background: Phytases are enzymes capable of degrading phytic acid and used in animal
feed supplementation in order to improve digestibility through the release of minerals such as
Objective: The main goal of this study was to express and characterize a Yersinia intermedia phytase
expressed in Escherichia coli cells.
Methods: The Y. intermedia phytase gene was synthesized and overexpressed in Escherichia coli
cells. The phytase recombinante (rPHY) was purified to homogeneity using a Ni-NTA column. The
biochemical and biophysical properties of the rPHY were measured in order to fully characterize
the recombinant enzyme. The following patents database were consulted: Espacenet, USPTO,
LATIPAT, Patent Scope, WIPO and Google Patents.
Results: The results showed that the rPHY is active at 37-40ºC and presented an optimal pH and
temperature of 8.0 and 40°C, respectively. The phytase rPHY was activated by Cu2+ ion and
showed resistance to trypsin and pepsin, retaining 55% of the activity at the ratio of 0.02. Furthermore,
the dissociation constant (Kd = 1.1150 ± 0.0087 mM), as estimated by a fluorescence binding
assay, suggests a medium affinity of the enzyme with the substrate.
Conclusion: The results of this article can be considered as innovative and for this reason, they
were protected by Intellectual Property Law in Brazil. Take together, the biochemical properties of
the rPHY could be useful in future for its industrial application of this enzyme as an additive in the