Hydrophobin (HFBI): A potential fusion partner for one-step purification of recombinant proteins from insect cells

Hydrophobins play an important role in binding and assembly of fungal surface structures as well as in medium–air interactions. These, hydrophobic properties provide interesting possibilities when purification of macromolecules is concerned. In aqueous micellar two-phase systems, based on surfactant...

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Bibliographic Details
Published in:Protein expression and purification 2008-05, Vol.59 (1), p.18-24
Main Authors: Lahtinen, Tomi, Linder, Markus B., Nakari-Setälä, Tiina, Oker-Blom, Christian
Format: Article
Language:English
Subjects:
Animals
Aqueous micellar two-phase system (AMTPS)
Avidin - biosynthesis
Avidin - isolation & purification
Baculoviridae
Baculovirus
Blotting, Western
Fluorescence scanning microscopy (FSM)
Fungal Proteins - chemistry
Fungal Proteins - genetics
Hydrophobin
Hypocrea jecorina
Micelles
Microscopy, Confocal
Protein purification
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - isolation & purification
Spodoptera
Surfactants
Trichoderma - chemistry
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Summary:Hydrophobins play an important role in binding and assembly of fungal surface structures as well as in medium–air interactions. These, hydrophobic properties provide interesting possibilities when purification of macromolecules is concerned. In aqueous micellar two-phase systems, based on surfactants, the water soluble hydrophobins are concentrated inside micellar structures and, thus, distributed to defined aqueous phases. This, one-step purification is attractive particularly when large-scale production of recombinant proteins is concerned. In the present study the hydrophobin HFBI of Trichoderma reesei was expressed as an N-terminal fusion with chicken avidin in baculovirus infected insect cells. The intracellular distribution of the recombinant fusion construct was analyzed by confocal microscopy and the protein subsequently purified from cytoplasmic extracts in an aqueous micellar two-phase system by using a non-ionic surfactant. The results show that hydrophobin and an avidin fusion thereof were efficiently expressed in insect cells and that these hydrophobic proteins could be efficiently purified from these cells in one-step by adopting an aqueous micellar two-phase system.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2007.12.014