Abstract
Cell surface display of proteins/peptides, in contrast to the conventional intracellular expression, has many attractive features. This arming technology is especially effective when yeasts are used as a host, because eukaryotic modifications that are often required for functional use can be added to the surface-displayed proteins/peptides. A part of various cell wall or plasma membrane proteins can be genetically fused to the proteins/peptides of interest to be displayed. This technology, leading to the generation of so-called “arming technology”, can be employed for basic and applied research purposes. In this article, we describe various strategies for the construction of arming yeasts, and outline the diverse applications of this technology to industrial processes such as biofuel and chemical productions, pollutant removal, and health-related processes, including oral vaccines. In addition, arming technology is suitable for protein engineering and directed evolution through high-throughput screening that is made possible by the feature that proteins/peptides displayed on cell surface can be directly analyzed using intact cells without concentration and purification. Actually, novel proteins/peptides with improved or developed functions have been created, and development of diagnostic/therapeutic antibodies are likely to benefit from this powerful approach.
Highlights
Arming technology in yeast using the cell surface display system is an innovative technology for the construction of whole-cell biocatalysts and protein engineering through high-throughput screening of protein libraries
Intracellular expression and extracellular secretion have been employed in conventional strategies for molecular breeding, the cell surface is an attractive location for heterologous gene expression
Surface-engineered cells prepared by cell cultivation are ready to use as microparticles covered by proteins/peptides, whereas troublesome protein purification and concentration procedures are required for the analysis and subsequent application of proteins produced by intracellular expression and extracellular secretion
Summary
Arming technology in yeast using the cell surface display system is an innovative technology for the construction of whole-cell biocatalysts and protein engineering through high-throughput screening of protein libraries. Cell surface-engineered yeasts constructed using the arming technology, have been termed “arming yeasts” [1,2,3]. Surface-engineered cells prepared by cell cultivation are ready to use as microparticles covered by proteins/peptides, whereas troublesome protein purification and concentration procedures are required for the analysis and subsequent application of proteins produced by intracellular expression and extracellular secretion. In the case of target proteins that have a high molecular mass or require glycosylation modification, yeasts are suitable hosts for cell surface display. We review the advances of molecular breeding of novel cells and protein engineering by arming technology in yeasts such as Saccharomyces cerevisiae, Pichia pastoris, and Yarrowia lipolytica
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