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Secretion of human interleukin-2 fused with green fluorescent protein in recombinatn pichia pastoris

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Abstract

Methylotrophic yeast Pichia pastoris is convenient for the expression of eukaryotic foreign proteins owing to its potential for posttranslational modifications, protein folding, and facile culturing. In this work, human interleukin (hIL)-2 was successfully produced as a secreted fusion form in recombinant P. pastoris. By employing green fluorescent protein (GFP) as a monitoring fusion partner, clear identification of fusion protein expression and quantification of intracellular hIL-2 were possible even though there was no correlation between culture supernatant fluorescence and secreted hIL-2 owing to high media interference. Importantly, by the addition of casamino acids in basal medium, we were able to enhance threefold amount of secreted hIL-2, which was present both as a fusion and as a clipped fragment.

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Authors and Affiliations

  1. Division of Molecular and Life Sciences & Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea

    Hyung Joon Cha

  2. Center for Biosystems Research & Department of Chemical Engineering, University of Maryland, 20742, College Park, Maryland

    Nimish N. Dalal & William E. Bentley

Authors
  1. Hyung Joon Cha
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  2. Nimish N. Dalal
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  3. William E. Bentley
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Correspondence to Hyung Joon Cha.

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Cha, H.J., Dalal, N.N. & Bentley, W.E. Secretion of human interleukin-2 fused with green fluorescent protein in recombinatn pichia pastoris . Appl Biochem Biotechnol 126, 1–11 (2005). https://doi.org/10.1007/s12010-005-0001-9

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  • DOI: https://doi.org/10.1007/s12010-005-0001-9

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