title: Hydrophobin (HFBI): A potential fusion partner for one-step purification of recombinant proteins from insect cells.
abstract: 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.
authors.affiliation: Department of Biological and Environmental Science, University of Jyväskylä, Nanoscience Center, PO Box 35, 40014 Jyväskylä, Finland.
authors.forename: Tomi
authors.initials: T
authors.lastname: Lahtinen
authors.forename: Markus B
authors.initials: MB
authors.lastname: Linder
authors.forename: Tiina
authors.initials: T
authors.lastname: Nakari-Setälä
authors.forename: Christian
authors.initials: C
authors.lastname: Oker-Blom
mesh_terms: D000818:Animals; D001360:Avidin / Q000096:biosynthesis / Q000302:isolation & purification*; D016367:Baculoviridae; D015153:Blotting, Western; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D008823:Micelles; D018613:Microscopy, Confocal; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D018411:Spodoptera; D014242:Trichoderma / Q000737:chemistry
chemical_list: D005656:Fungal Proteins; D008823:Micelles; D011993:Recombinant Fusion Proteins; D001360:Avidin