title: Bioseparation of recombinant proteins from plant extract with hydrophobin fusion technology.
abstract: Two main hurdles hinder the widespread acceptance of plants as a preferred protein expression platform: low accumulation levels and expensive chromatographic purification methods. Fusion of proteins of interest to fungal hydrophobins has provided a tool to address both accumulation and purification issues. In this method, we describe the one-step purification of a GFP-HFBI fusion from crude plant extract using an aqueous two-phase system (ATPS). ATPS can be carried out in a very short time frame, yields relatively pure protein with very few contaminants, and does not require any chromatographic column steps. This purification system takes advantage of the affinity of hydrophobins to the micellar phase of widely available nonionic surfactants, such as Triton X-114, and can be easily scaled up for industrial-scale protein purification.
authors.affiliation: VTT Technical Research Centre of Finland, Tietotie, Finland. jussi.joensuu@vtt.fi
authors.forename: Jussi J
authors.initials: JJ
authors.lastname: Joensuu
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Conley
authors.forename: Markus B
authors.initials: MB
authors.lastname: Linder
authors.forename: Rima
authors.initials: R
authors.lastname: Menassa
mesh_terms: D019149:Bioreactors; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism; D008823:Micelles; D018613:Microscopy, Confocal; D017830:Octoxynol; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry*; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D013501:Surface-Active Agents / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry*; D005466:Fluorocarbons
chemical_list: D005656:Fungal Proteins; D049452:Green Fluorescent Proteins; D007093:Imidazoles; D008823:Micelles; D017830:Octoxynol; D010936:Plant Extracts; D011092:Polyethylene Glycols; D011994:Recombinant Proteins; D013501:Surface-Active Agents; C046634:1-(heptafluorobutyryl)imidazole; D005466:Fluorocarbons; C010615:Nonidet P-40