title: Cloning and expression of Aspergillus flavus urate oxidase in Pichia pastoris.
abstract: Urate oxidase is an important enzyme with therapeutic and diagnostic applications. Rasburicase is a recombinant urate oxidase enzyme approved by FDA to use in the treatment of hyperuricemia conditions. Various hosts such as Saccharomyces cerevisiae, Hansenula polymorpha and Escherichia coli have been used to express the enzyme. Today, Pichia pastoris is considered as an important host for heterologous protein expression since it has beneficial characteristics such as strong promoters, simple scale up, post translational modifications, high cell density cultivation and simple genetic manipulation. In this study, Aspergillus flavus urate oxidase gene was cloned in pPICZαA expression vector and expressed in P. pastoris. The recombinant urate oxidase was expressed in secretory form and was confirmed through RT-PCR, SDS-PAGE analysis and western blotting. The enzyme activity was determined using a colorimetric assay. A production yield of 0.43 U/ml of culture supernatant was obtained.
authors.affiliation: Department of Biotechnology, Faculty of Science, University of Tehran, 14174 Tehran, Iran.
authors.forename: Ramin
authors.initials: R
authors.lastname: Fazel
authors.affiliation: Medical Biotechnology department, Fungal Biotechnology group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Najmeh
authors.initials: N
authors.lastname: Zarei
authors.affiliation: Department of Biotechnology, Faculty of Science, University of Tehran, 14174 Tehran, Iran.
authors.forename: Nasser
authors.initials: N
authors.lastname: Ghaemi
authors.affiliation: Pharmaceurical Biotechnology department, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Namvaran
authors.affiliation: Medical Biotechnology department, Fungal Biotechnology group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Somayeh
authors.initials: S
authors.lastname: Enayati
authors.affiliation: Medical Biotechnology department, Fungal Biotechnology group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Esmat
authors.initials: E
authors.lastname: Mirabzadeh Ardakani
authors.affiliation: Medical Biotechnology department, Fungal Biotechnology group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Azizi
authors.affiliation: Medical Biotechnology department, Fungal Biotechnology group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Khalaj
keywords: Pichia; Rasburicase; Urate oxidase
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