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Aqueous two-phase systems containing urea: Influence of protein structure on protein partitioning

✍ Scribed by Christian Rämsch; Lutz B. Kleinelanghorst; Esther A. Knieps; Jörg Thömmes; Maria-Regina Kula

Publisher
John Wiley and Sons
Year
2000
Tongue
English
Weight
200 KB
Volume
69
Category
Article
ISSN
0006-3592

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✦ Synopsis

During recombinant E. coli fermentation with high-expression levels inclusion bodies are often formed. Aqueous two-phase systems have been successfully used in the presence of urea for the initial recovery step of inclusion bodies from E. coli. Basic studies of the complex interactions are lacking. For a systematic study of protein partitioning in the presence of urea we selected T4-lysozyme mutants with different thermal stability as a model. The stabilization of these variants by phase components was investigated measuring the fluorescence emission of tryptophan residues in the protein. Protein structure was stabilized at pH 7 in the order of S0(4)(2-) >> PEG = Dextran > H(2)O. The conformation of proteins was shown to have a strong influence on the partitioning in aqueous two-phase systems. Tryptophan and its homologuous di- and tripeptdides were partitioned in similar phase systems to normalize for contribution from hydrophobic interactions.

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