title: Expression and characterization of human interferon-beta1 in the methylotrophic yeast Pichia pastoris.
abstract: We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHuIFN-beta1) was secreted from shake-flask-grown P. pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.u. (international units)/ml (6-12 mg/litre). An rHuIFN-beta1 with an N-terminal sequence identical with that of native HuIFN-beta1 was purified and the specific activity was determined (2-3 x 10(7) i.u./mg). It was found that the secreted recombinant protein was partially N-glycosylated.
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444a, 11001 Belgrade, Serbia and Montenegro. nataskoko@yahoo.it
authors.forename: Natasa
authors.initials: N
authors.lastname: Skoko
authors.forename: Barbara
authors.initials: B
authors.lastname: Argamante
authors.forename: Natasa Kovacević
authors.initials: NK
authors.lastname: Grujicić
authors.forename: Sergio Gabriel
authors.initials: SG
authors.lastname: Tisminetzky
authors.forename: Vladimir
authors.initials: V
authors.lastname: Glisin
authors.forename: Goran
authors.initials: G
authors.lastname: Ljubijankić
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D006801:Humans; D016899:Interferon-beta / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014162:Transfection / Q000379:methods; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects
chemical_list: D011994:Recombinant Proteins; D016899:Interferon-beta