title: High-level expression and purification of recombinant human catalase in Pichia pastoris.
abstract: 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.
authors.affiliation: Department of Drug Biosynthesis, School of Pharmacy, Fudan University, 138 Yi Xue Yuan Road, Shanghai 200032, China. shixunlong@yahoo.com.cn
authors.forename: Xun-Long
authors.initials: XL
authors.lastname: Shi
authors.forename: Mei-Qing
authors.initials: MQ
authors.lastname: Feng
authors.forename: Jian
authors.initials: J
authors.lastname: Shi
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Shi
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhong
authors.forename: Pei
authors.initials: P
authors.lastname: Zhou
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D002374:Catalase