title: Characterization of recombinant fungal phytase (phyA) expressed in tobacco leaves.
abstract: The phyA gene from Aspergillus ficuum coding for a 441-amino-acid full-length phytase was expressed in Nicotiana tabacum (tobacco) leaves. The expressed phytase was purified to homogeneity using ion-exchange column chromatography. The purified phytase was characterized biochemically and its kinetic parameters were determined. When the recombinant phytase was compared with its counterpart from Aspergillus ficuum for physical and enzymatic properties, it was found that catalytically the recombinant protein was indistinguishable from the native phytase. Except for a decrease in molecular mass, the overexpressed recombinant phytase was virtually the same as the native fungal phytase. While the temperature optima of the recombinant protein remain unchanged, the pH optima shifted from pH 5 to 4. The results are encouraging enough to open the possibility of overexpressing phyA gene from Aspergillus ficuum in other crop plants as an alternative means of commercial production of this important enzyme.
authors.affiliation: Southern Regional Research Center, ARS, USDA, New Orleans, Louisiana, 70124, USA.
authors.forename: A H
authors.initials: AH
authors.lastname: Ullah
authors.forename: K
authors.initials: K
authors.lastname: Sethumadhavan
authors.forename: E J
authors.initials: EJ
authors.lastname: Mullaney
authors.forename: T
authors.initials: T
authors.lastname: Ziegelhoffer
authors.forename: S
authors.initials: S
authors.lastname: Austin-Phillips
mesh_terms: D010832:6-Phytase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003362:Cost-Benefit Analysis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006031:Glycosylation; D007700:Kinetics; D008970:Molecular Weight; D010658:Phenylglyoxal / Q000494:pharmacology; D018515:Plant Leaves / Q000378:metabolism; D010947:Plants, Toxic; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D010832:6-Phytase; D010658:Phenylglyoxal