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Expression and characterization of human interferon‐β1 in the methylotrophic yeast Pichia pastoris
Author(s) -
Skoko Nataša,
Argamante Barbara,
Grujičić Nataša Kovačević,
Tisminetzky Sergio Gabriel,
Glišin Vladimir,
Ljubijankić Goran
Publication year - 2003
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20030065
Subject(s) - pichia pastoris , heterologous , recombinant dna , yeast , heterologous expression , saccharomyces cerevisiae , pichia , biochemistry , biology , alpha interferon , microbiology and biotechnology , biological activity , interferon , gene , in vitro , virology
We describe the heterologous expression of a human interferon‐β1 in the methylotrophic yeast Pichia pastoris . Biologically active recombinant human interferon‐β1 (rHuIFN‐β1) was secreted from shake‐flask‐grown P. pastoris cells into the medium using the Saccharomyces cerevisiae α‐mating factor prepro‐leader sequence at the level of (1–3)×10 5 i.u. (international units)/ml (6–12 mg/litre). An rHuIFN‐β1 with an N‐terminal sequence identical with that of native HuIFN‐β1 was purified and the specific activity was determined (2–3×10 7 i.u./mg). It was found that the secreted recombinant protein was partially N‐glycosylated.