[Construction of a cell-surface expression system in Trichoderma reesei].

Area of Science:

• Biotechnology
• Mycology
• Molecular Biology

Background:

• The glycosylphosphatidylinositol (GPI) anchored cell wall protein (GPI-CWP) AfMp1p from Aspergillus fumigatus possesses a C-terminal signal for cell wall localization.
• Trichoderma reesei is a Generally Recognized As Safe (GRAS) fungus with significant industrial applications.
• Developing cell-surface expression systems in T. reesei is of industrial interest for protein production.

Purpose of the Study:

• To investigate the functionality of the GPI signal from A. fumigatus AfMp1p in Trichoderma reesei.
• To establish a heterologous cell-surface expression system in T. reesei using the A. fumigatus GPI signal.

Main Methods:

• The GPI signal sequence from A. fumigatus AfMp1p was genetically fused to the C-terminus of green fluorescent protein (GFP).
• The resulting fusion construct was transformed into T. reesei.
• Optimization of T. reesei transformation protocols and detailed analysis of GFP expression profiles were performed.
• Cellular localization of the GFP fusion protein was experimentally determined.

Main Results:

• Fluorescent imaging confirmed the presence of the GFP fusion protein on the cell surface.
• Western blot analysis corroborated the cell-wall localization of the expressed GFP fusion protein in T. reesei.
• The results demonstrate successful targeting of the protein to the cell wall.

Conclusions:

• The GPI signal from Aspergillus fumigatus is effectively recognized and functional within Trichoderma reesei.
• The developed expression system allows for the expression and localization of heterologous proteins to the cell wall of T. reesei.
• This system holds potential for biotechnological applications requiring cell-wall displayed proteins in T. reesei.