Expression of a fungal hydrophobin in the Saccharomyces cerevisiae cell wall: effect on cell surface properties and immobilization.
The aim of this work was to modify the cell surface properties of Saccharomyces cerevisiae by expression of the HFBI hydrophobin of the filamentous fungus Trichoderma reesei on the yeast cell surface. The second aim was to study the immobilization capacity of the modified cells. Fusion to the Flo1p flocculin was used to target the HFBI moiety to the cell wall. Determination of cell surface characteristics with contact angle and zeta potential measurements indicated that HFBI-producing cells are more apolar and slightly less negatively charged than the parent cells. Adsorption of the yeast cells to different commercial supports was studied. A twofold increase in the binding affinity of the hydrophobin-producing yeast to hydrophobic silicone-based materials was observed, while no improvement in the interaction with hydrophilic carriers could be seen compared to that of the parent cells. Hydrophobic interactions between the yeast cells and the support are suggested to play a major role in attachment. Also, a slight increase in the initial adsorption rate of the hydrophobin yeast was observed. Furthermore, due to the engineered cell surface, hydrophobin-producing yeast cells were efficiently separated in an aqueous two-phase system by using a nonionic polyoxyethylene detergent, C(12-18)EO(5).
Nakari-Setälä
Tiina
T
VTT Biotechnology, PO Box 1500, FIN-02044 VTT, Finland. tiina.nakari-setala@vtt.fi
Azeredo
Joana
J
Henriques
Mariana
M
Oliveira
Rosário
R
Teixeira
José
J
Linder
Markus
M
Penttilä
Merja
M
D001665:Binding Sites; D002473:Cell Wall / Q000378:metabolism*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins; D014242:Trichoderma / Q000235:genetics
C087578:FLO1 protein, S cerevisiae; D005656:Fungal Proteins; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins