title: Purification and characterization of a thermostable extracellular phytase from Bacillus licheniformis PFBL-03.
abstract: Extracellular phytase from Bacillus licheniformis PFBL-03 was purified in three steps by using ammonium sulfate precipitation, ion-exchange chromatography on a DEAE Sephadex A-50 column, and gel filtration chromatography on Sephadex G-100. The single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested that the enzyme was homogeneous. The molecular mass determined from SDS-PAGE was 36 kD. The enzyme yield was 10% while the purification fold was 39. The purified phytase exhibited optimum activity at 55°C and was able to retain 55% of its original activity after 60 min of incubation at 80°C. The purified enzyme had optimum pH of 6.0 and was stable over a pH range of 4.0 to 7.5. The kinetic parameters K(m) and V(max) of the purified phytase for sodium phytate were 4.7 mM and 49.01 µmol/min. The activity of the enzyme was enhanced in the presence of Ca²⁺ but completely inhibited by Fe²⁺, Mn²⁺, and Cu²⁺. The exhibited characteristics of the purified phytase from Bacillus licheniformis PFBL-03 show that the enzyme has potential for applications in the animal feed industry.
authors.affiliation: a Department of Biochemistry , Federal University of Technology , Akure , Nigeria.
authors.forename: Feyisola O
authors.initials: FO
authors.lastname: Fasimoye
authors.forename: Folasade M
authors.initials: FM
authors.lastname: Olajuyigbe
authors.forename: Morakinyo D
authors.initials: MD
authors.lastname: Sanni
mesh_terms: D010832:6-Phytase / Q000302:isolation & purification* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D007700:Kinetics
chemical_list: D010832:6-Phytase