Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications

Abstract

Yeast surface display (YSD) is a “whole-cell” platform used for the heterologous expression of proteins immobilized on the yeast’s cell surface. YSD combines the advantages eukaryotic systems offer such as post-translational modifications, correct folding and glycosylation of proteins, with ease of cell culturing and genetic manipulation, and allows of protein immobilization and recovery. Additionally, proteins displayed on the surface of yeast cells may show enhanced stability against changes in temperature, pH, organic solvents, and proteases. This platform has been used to study protein-protein interactions, antibody design and protein engineering. Other applications for YSD include library screening, whole-proteome studies, bioremediation, vaccine and antibiotics development, production of biosensors, ethanol production and biocatalysis. YSD is a promising technology that is not yet optimized for biotechnological applications. This mini review is focused on recent strategies to improve the efficiency and selection of displayed proteins. YSD is presented as a cutting-edge technology for the vectorial expression of proteins and peptides. Finally, recent biotechnological applications are summarized. The different approaches described herein could allow for a better strategy cascade for increasing protein/peptide interaction and production.

Bottom-Up Design: A Modular Golden Gate Assembly Platform of Yeast Plasmids for Simultaneous Secretion and Surface Display of Distinct FAP Fusion Proteins

“…For the TPD–rFAP secretion reporter, we created three modules: secretion leader, TPD, and the dL5 FAP to allow easy exchange of the leader and TPD and be capable of modulating TPD protein secretion levels, if needed. In yeast secretion, TPDs can be strongly dependent on the type of secretion leader and TPD, and therefore, it is crucial to have the flexibility to change the secretion leaders or modify TPDs. For the NB–gFAP display reporter, we created four modules: Aga2p-linker-TPD barcode; NB; Sag1p; and AM2.2 or ScFv1 gFAP.…”

“…cerevisiae, GG has been mainly used for engineering biosynthetic pathways − and remodeling the genome, often using CRISPR technology to exploit efficient yeast homologous recombination. , GG is also useful for yeast display, , a versatile technology where yeast-expressed proteins of interest are displayed on the cell surface as genetic fusions to cell wall proteins. Yeast-displayed proteins may serve as extracellular enzymes or as candidate affinity reagents …”

Taking a Breather: Advances in Interleukin 5 Inhibition for Asthma Relief

“…Akin to the concepts of phage display, yeast display sees antibody libraries being bound to yest cells via fusion with the N terminus of the Aga2p subunit [ 36 ]. In vitro surface display technology holds a specific advantage over ex vivo technologies, that is the incorporation of protein expression tags allowing for the selection of correctly assembled antibodies [ 37 ]. Another advantage specific to cell-based display technologies is the ability to use flow cytometry to analyse each cell independently.…”

Saccharomyces cerevisiae surface display of endolysin LysKB317 for control of bacterial contamination in corn ethanol fermentations

“…The use of a yeast surface display platform to anchor recombinant proteins to the cell wall of yeast has been widely adapted in the biomedical eld for antibody and antimicrobial peptide engineering, protein stability, protein-protein interaction, and enzyme metabolic studies [21][22][23][24][25][26][27][28][29]. A common yeast display system involves fusing recombinant proteins to the α-agglutinin mating protein Aga2p and expressing extracellularly in yeast that secrete the β 1,6 glucan-anchored Aga1p domain of α-agglutinin to form an Aga1p-Aga2p complex via two disul de bonds [21,30].…”

“…In this platform, epitope detection can be performed with the protein of interest when correctly expressed, folded and surface displayed. In addition, protein aggregation in the cytoplasm is reduced and the need for protein puri cation in the downstream process is eliminated with the yeast surface display platform [24]. Recently, Chun et al, demonstrated antibacterial activity against Staphylococcus aureus using surface displayed expression of S. aureus phage endolysin LysSA11 on S. cerevisiae [31].…”