초록
<P><B>Background</B></P><P>Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) is a glycoprotein that has been approved by the FDA for the treatment of neutropenia and leukemia in combination with chemotherapies. Recombinant hGM-CSF is produced industrially using the baker’s yeast, <I>Saccharomyces cerevisiae</I>, by large-scale fermentation. The methylotrophic yeast, <I>Pichia pastoris</I>, has emerged as an alternative host cell system due to its shorter and less immunogenic glycosylation pattern together with higher cell density growth and higher secreted protein yield than <I>S. cerevisiae</I>. In this study, we compared the pipeline from gene to recombinant protein in these two yeasts.</P><P><B>Results</B></P><P>Codon optimization <I>in silico</I> for both yeast species showed no difference in frequent codon usage. However, rhGM-CSF expressed from <I>S. cerevisiae</I> BY4742 showed a significant discrepancy in molecular weight from those of <I>P. pastoris</I> X33. Analysis showed purified rhGM-CSF species with molecular weights ranging from 30 to more than 60 kDa. Fed-batch fermentation over 72 h showed that rhGM-CSF was more highly secreted from <I>P. pastoris</I> than <I>S. cerevisiae</I> (285 and 64 mg total secreted protein/L, respectively). Ion exchange chromatography gave higher purity and recovery than hydrophobic interaction chromatography. Purified rhGM-CSF from <I>P. pastoris</I> was 327 times more potent than rhGM-CSF from <I>S. cerevisiae</I> in terms of proliferative stimulating capacity on the hGM-CSF-dependent cell line, TF-1.</P><P><B>Conclusion</B></P><P>Our data support a view that the methylotrophic yeast <I>P. pastoris</I> is an effective recombinant host for heterologous rhGM-CSF production.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1186/s13104-017-2471-6) contains supplementary material, which is available to authorized users.</P>