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

85 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

Fingerprint

Recombinant Proteins
Surface-Active Agents
Purification
Proteins
Recombinant Fusion Proteins
Disulfides
Nonionic surfactants
Fungi
Membrane Proteins
Phase separation
Fusion reactions
Amino Acids
Temperature

Keywords

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

Cite this

Linder, Markus ; Qiao, Mingqiang ; Laumen, Frank ; Selber, Klaus ; Hyytiä, Teppo ; Nakari-Setälä, Tiina ; Penttilä, Merja. / Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems. In: Biochemistry. 2004 ; Vol. 43, No. 37. pp. 11873-11882.
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Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems. / Linder, Markus; Qiao, Mingqiang; Laumen, Frank; Selber, Klaus; Hyytiä, Teppo; Nakari-Setälä, Tiina; Penttilä, Merja.

In: Biochemistry, Vol. 43, No. 37, 2004, p. 11873-11882.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems

AU - Linder, Markus

AU - Qiao, Mingqiang

AU - Laumen, Frank

AU - Selber, Klaus

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AU - Nakari-Setälä, Tiina

AU - Penttilä, Merja

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AB - 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.

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