Yeast arming systems: pros and cons of different protein anchors and other elements required for display

Abstract

Yeast display is a powerful strategy that consists in exposing peptides or proteins of interest on the cell surface of this microorganism. Ever since initial experiments with this methodology were carried out, its scope has extended and many applications have been successfully developed in different science and technology fields. Several yeast display systems have been designed, which all involve introducting into yeast cells the gene fusions that contain the coding regions of a signal peptide, an anchor protein, to properly attach the target to the cell surface, and the protein of interest to be exposed, all of which are controlled by a strong promoter. In this work, we report the description of such elements for the alternative systems introduced by focusing particularly on anchor proteins. The comparisons made between them are included whenever possible, and the main advantages and inconveniences of each one are discussed. Despite the huge number of publications on yeast surface display and the revisions published to date, this topic has not yet been widely considered. Finally, given the growing interest in developing systems for non-Saccharomyces yeasts, the main strategies reported for some are also summarized.

Evolving a Peptide: Library Platforms and Diversification Strategies

“…Examples of surface display libraries include cysteine knot peptides (knottins) [105] and lanthipeptides [106], whereas head-to-tail cyclized peptide libraries [96,107] (see the SICLOPPS method description below) are expressed intracellularly. The main advantage of yeast display is its eukaryotic protein expression mechanism, which allows for complex post-translational modifications, and quantitative library screening through FACS [108,109]. Disadvantages include smaller library sizes due to low transformation efficiency [110], and lower affinity caused by unintended multivalent binding to oligomeric targets, although this can be surmounted by applying kinetic selections [111].…”

Pichia pastoris as a Versatile Cell Factory for the Production of Industrial Enzymes and Chemicals: Current Status and Future Perspectives

“…This hypothesis is supported by several experimental studies. With the same anchor system (α‐agglutinin), 9 × 10 5 molecules of enzyme per cell was determined for P. pastoris where the value ranged from 1.5 × 10 5 to 2.75 × 10 5 for S. cerevisiae . Display of lipase on P. pastoris has been extensively reported for biodiesel production (refer to Yan et al for a detailed review) or synthesis of various kind of esters .…”

Molecular evolution of peptides by yeast surface display technology

“…[14][15][16] In addition to tuning the affinity and specificity of multiple proteins and peptides towards a wide range of targets, yeast surface display technology has also been successfully used for epitope mapping, to improve the recombinant production and the stability of the molecules of interest as well as to engineer the function of several enzymes. [14][15][16] Although diverse yeast strains and various cell wall anchors have been used to display a large variety of protein and peptide scaffolds, [17][18][19] the Saccharomyces cerevisiae Aga1-Aga2 display system remains the most commonly used. In this arrangement, the molecule of interest is expressed as fusion to the Aga2 protein that is linked to the membrane anchored a-agglutinin Aga1 protein through two disulfide bridges, resulting in a covalent complex on the surface of the yeast cell.…”