A Highly Efficient Indirect P. pastoris Surface Display Method Based on the CL7/Im7 Ultra-High-Affinity System

Area of Science:

• Biotechnology
• Synthetic Biology
• Protein Engineering

Background:

• Cell surface display systems are crucial for applications like vaccine generation and bio-conversion.
• Traditional yeast surface display methods face challenges in efficiency and universality.
• Developing robust and efficient display platforms is essential for advancing biomolecule applications.

Purpose of the Study:

• To develop a highly efficient and universal indirect yeast surface display system.
• To demonstrate the capability of the system for displaying various target proteins.
• To establish a robust platform for biomolecule immobilization on Pichia pastoris.

Main Methods:

• An indirect display strategy was employed using Pichia pastoris.
• Im7 proteins were anchored to the P. pastoris cell surface.
• Target proteins were fused with a CL7 tag for high-affinity binding to Im7.

Main Results:

• Highly efficient display of target proteins, including fluorescent proteins (sfGFP, mCherry) and enzymes (human Arginase I), was achieved.
• The system leverages the ultra-high-affinity interaction between Im7 and CL7 for robust immobilization.
• The indirect P. pastoris surface display approach proved to be highly efficient.

Conclusions:

• The developed indirect P. pastoris surface display system offers a highly efficient method for biomolecule immobilization.
• This platform provides a robust and versatile tool for protein engineering, vaccine development, and other biotechnological applications.
• The Im7-CL7 interaction enables reliable and high-affinity display of diverse proteins.