Skoko Natasa, Argamante Barbara, Grujicić Natasa Kovacević, Tisminetzky Sergio Gabriel, Glisin Vladimir, Ljubijankić Goran
Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 444a, 11001 Belgrade, Serbia and Montenegro.
Biotechnol Appl Biochem. 2003 Dec;38(Pt 3):257-65. doi: 10.1042/BA20030065.
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.
我们描述了人干扰素-β1在甲基营养型酵母毕赤酵母中的异源表达。具有生物活性的重组人干扰素-β1（rHuIFN-β1）利用酿酒酵母α-交配因子前原导序列，从摇瓶培养的毕赤酵母细胞分泌到培养基中，水平为（1-3）×10⁵国际单位/毫升（6-12毫克/升）。纯化了一种N端序列与天然HuIFN-β1相同的rHuIFN-β1，并测定了其比活性（2-3×10⁷国际单位/毫克）。发现分泌的重组蛋白部分进行了N-糖基化。
