High-level expression and purification of recombinant human catalase in Pichia pastoris.
Catalase is one of the antioxidant enzymes and is involved in many pathophysiologic processes and human diseases. This study focused on high-level expression and purification of recombinant catalase in Pichia pastoris. The cDNA encoding catalase was cloned by RT-PCR from Fetal liver of Homo sapiens. After PCR and construction of expression vector pPIC9K-CAT, human catalase was expressed highly in P. pastoris yeast SMD1168 and secreted into the culture medium. The secreted catalase was purified to a purity of 95% by ammonium sulfate fractionation, anionic exchange-chromatography, and Macro-prep Ceramic Hydroxyapatite with a overall yield of 60%. This study provides a new method for large-scale expression and purification of recombinant protein catalase.
Shi
Xun-Long
XL
Department of Drug Biosynthesis, School of Pharmacy, Fudan University, 138 Yi Xue Yuan Road, Shanghai 200032, China. shixunlong@yahoo.com.cn
Feng
Mei-Qing
MQ
Shi
Jian
J
Shi
Zhi-Hui
ZH
Zhong
Jiang
J
Zhou
Pei
P
D002374:Catalase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007218:Industrial Microbiology / Q000379:methods*; D010843:Pichia / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D014170:Transformation, Genetic
D018076:DNA, Complementary; D011994:Recombinant Proteins; D002374:Catalase