Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems

Markus Linder, Mingqiang Qiao, Frank Laumen, Klaus Selber, Teppo Hyytiä, Tiina Nakari-Setälä, Merja Penttilä

    Research output: Contribution to journalArticleScientificpeer-review

    94 Citations (Scopus)

    Abstract

    In this work we describe the new concept of using fungal hydrophobins as efficient tags for purification of recombinant fusion proteins by aqueous two-phase separation. Hydrophobins are a group of small surface-active proteins produced by filamentous fungi. Some characteristics of hydrophobins are that they are relatively small (approximately 100 amino acids), they contain eight disulfide-forming Cys residues in a conserved pattern, and they self-assemble on interfaces. The aqueous two-phase systems studied were based on nonionic surfactants that phase-separate at certain temperatures. We show that the use of hydrophobins as tags has many advantages such as high selectivity and good yield and is technically very simple to perform. Fusion proteins with target proteins of different molecular size were compared to the corresponding free proteins using a set of different surfactants. This gave an understanding on which factors influence the separation and what rationale should be used for optimization. This unusually strong and specific interaction between polymeric surfactants and a soluble protein shows promise for new developments in interfacing proteins and nonbiological materials for other applications as well.
    Original languageEnglish
    Pages (from-to)11873-11882
    JournalBiochemistry
    Volume43
    Issue number37
    DOIs
    Publication statusPublished - 2004
    MoE publication typeA1 Journal article-refereed

    Keywords

    • proteins
    • recombinant proteins
    • hydrophobins
    • filamentous fungi
    • fusion proteins
    • tags

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