Cloning and expression of Aspergillus flavus urate oxidase in Pichia pastoris

被引:24
|
作者
Fazel, Ramin [1 ]
Zarei, Najmeh [2 ]
Ghaemi, Nasser [1 ]
Namvaran, Mohammad Mehdi [3 ]
Enayati, Somayeh [2 ]
Ardakani, Esmat Mirabzadeh [2 ]
Azizi, Mohammad [2 ]
Khalaj, Vahid [2 ]
机构
[1] Univ Tehran, Fac Sci, Dept Biotechnol, Tehran 14174, Iran
[2] Pasteur Inst Iran, Biotechnol Res Ctr, Fungal Biotechnol Grp, Med Biotechnol Dept, Tehran, Iran
[3] Shahid Beheshti Univ Med Sci, Pharmaceur Biotechnol Dept, Tehran, Iran
来源
SPRINGERPLUS | 2014年 / 3卷
关键词
Urate oxidase; Pichia; Rasburicase; ESCHERICHIA-COLI; HANSENULA-POLYMORPHA; PROTEIN-PRODUCTION; URIC-ACID; GENE; HYPERURICEMIA; RASBURICASE; SYSTEMS;
D O I
10.1186/2193-1801-3-395
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 pPICZaA 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.
引用
收藏
页码:1 / 7
页数:7
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