Construction of a cell-surface expression system in Trichoderma reesei

Abstract:

Abstract:［Objective］AfMp1p is a glycosylphosphatidylinositol (GPI) anchored cell wall protein (GPI-CWP) identified in filamentous fungus Aspergillus fumigatus，which contains a specific C terminal signal for cell wall localization．Trichoderma reesei is known as a safe fungal species (GRAS) and widely used in the industry．Thus，developing of the cell-surface expression systems in T．reesei is of industrial interest．［Methods］The GPI signal from the A．fumigatus AfMp1p was fused to the C-terminal of green fluorescent protein (GFP) and transformed into T．reesei． The optimization of T．reesei transformation and the expression profiles of the GFP were investigated in detail．The cellular location of the GFP fusion protein was detected．［Results］Fluorescent image analysis and Western blot analysis indicate that the GFP fusion protein locates on the cell wall of T．reesei．［Conclusion］According to these results，the GPI signal from A．fumigatus can be recognized in T．reesei and the expression system constructed in this study can be used to express heterogeneous protein in the cell wall of T．reesei.