Development of novel surface display platforms for anchoring heterologous proteins in Saccharomyces cerevisiae

Abstract

Background Cell surface display of recombinant proteins has become a powerful tool for biotechnology and biomedical applications. As a model eukaryotic microorganism, Saccharomyces cerevisiae is an ideal candidate for surface display of heterologous proteins. However, the frequently used commercial yeast surface display system, the a-agglutinin anchor system, often results in low display efficiency. Results We initially reconstructed the a-agglutinin system by replacing two anchor proteins with one anchor protein. By directly fusing the target protein to the N-terminus of Aga1p and inserting a flexible linker, the display efficiency almost doubled, and the activity of reporter protein α-galactosidase increased by 39%. We also developed new surface display systems. Six glycosylphosphatidylinositol (GPI) anchored cell wall proteins were selected to construct the display systems. Among them, Dan4p and Sed1p showed higher display efficiency than the a-agglutinin anchor system. Linkers were also inserted to eliminate the effects of GPI fusion on the activity of the target protein. We further used the newly developed Aga1p, Dan4p systems and Sed1p system to display exoglucanase and a relatively large protein β-glucosidase, and found that Aga1p and Dan4p were more suitable for immobilizing large proteins. Conclusion Our study developed novel efficient yeast surface display systems, that will be attractive tools for biotechnological and biomedical applications Electronic supplementary material The online version of this article (10.1186/s12934-019-1133-x) contains supplementary material, which is available to authorized users.

Surface display system for probiotics and its application in aquaculture

“…2019). Yang et al (2019) reconstructed the a‐agglutinin system by replacing two anchor proteins with one anchor protein and found that by directly fusing the target protein to the N‐terminus of the Aga1 protein and inserting a flexible linker, the display efficiency almost doubled. Mathew et al (2018) anchored HIV envelope protein Env to the yeast cell wall in which Env was either attached directly to the yeast cell wall α‐agglutinin as a C‐terminal fusion or fused to the secreted protein Aga2p either as an N‐ or C‐ terminal fusion, and found fusion of Aga2p at the C‐terminal provided higher expression levels by flow cytometry (Mathew et al .…”

Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications

“…In yeast, typically, this comprises a cell wall protein (CWP) linked to glycosylphosphatidylinositol (GPI) ( Tanaka and Kondo, 2015 ; Lim et al, 2017 ). This platform combines fine-tuned gene expression and protein immobilization, which simplifies the purification process and allows the reuse and recovery of biocatalysts ( Yuzbasheva et al, 2011 ; Liu et al, 2014 ; Yang et al, 2019 ).…”

Surface Display—An Alternative to Classic Enzyme Immobilization

“…This approach was used for surface display of Trypanosoma cruzi trans-sialidase [25], Aspergillus niger lipase ANL [26], mammalian membrane protein CD47 [27], and a number of mammalian antibodies and receptors [29,49,50]. Yang et al [51] constructed α-galactosidase fusion to the N-terminus of Aga1 with a flexible linker of 17 amino acids (composed of Ser and Gly) inserted between Aga1 part and α-galactosidase, and accomplished almost doubled display efficiency and 39% activity incensement in comparison to construct in which α-galactosidase was fused to the N-terminus of Aga1 without the linker. Tang et al [52] used an Aga1 and Aga2 protein pair display system for the construction of synthetic cellulosomes.…”

“…Using the Sed1 promoter was also favorable because efficient heterologous protein display was achieved by growth fa or prolonged period without any changes in the carbon source. Yang et al [51] constructed and compared fusions of α-galactosidase with serine-and threonine-rich regions and the GPI binding domains of Sed1, Cwp2, Dan4, Tos6, Srp2, and Pry3. Among them, Dan4 and Sed1 showed the highest display efficiency, but display efficiency and enzyme activities were not entirely dependable.…”