title: Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology.
abstract: Insufficient accumulation and the lack of efficient purification methods are the two major bottlenecks hindering the recombinant production of many proteins. Alternative production schemes are urgently needed for proteins that remain challenging to express and purify with conventional techniques. We have found that hydrophobin fusions targeted to endoplasmic reticulum (ER) can enhance the expression of target proteins simultaneously providing means for straightforward purification. Here we show that hydrophobin fusion technology induces formation of large protein bodies in the filamentous fungus Trichoderma reesei. The fusion protein remained soluble in the ER-derived protein bodies. A simple and scalable aqueous two-phase system was demonstrated to purify the hydrophobin fusion protein GFP-HFBI from the complex intracellular extracts with a recovery of up to 62%.
authors.affiliation: VTT Biotechnology, VTT Technical Research Centre of Finland, Espoo 02044 VTT, Finland.
authors.forename: Eero
authors.initials: E
authors.lastname: Mustalahti
authors.forename: Markku
authors.initials: M
authors.lastname: Saloheimo
authors.forename: Jussi Joonas
authors.initials: JJ
authors.lastname: Joensuu
mesh_terms: D018533:Biomass; D019149:Bioreactors / Q000382:microbiology; D001709:Biotechnology / Q000379:methods*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014242:Trichoderma / Q000254:growth & development / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D043366:Cellulose 1,4-beta-Cellobiosidase