Secretion of human interleukin-2 fused with green fluorescent protein in recombinant Pichia pastoris.
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.
Cha
Hyung Joon
HJ
Division of Molecular and Life Sciences, Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea. hjcha@postech.ac.kr
Dalal
Nimish N
NN
Bentley
William E
WE
D024861:Genetic Enhancement / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins