Surface Display—An Alternative to Classic Enzyme Immobilization

Abstract

Enzyme immobilization to solid matrices often presents a challenge due to protein conformation sensitivity, desired enzyme purity, and requirements for the particular carrier properties and immobilization technique. Surface display of enzymes at the cell walls of microorganisms presents an alternative that has been the focus of many research groups worldwide in different fields, such as biotechnology, energetics, pharmacology, medicine, and food technology. The range of systems by which a heterologous protein can be displayed at the cell surface allows the appropriate one to be found for almost every case. However, the efficiency of display systems is still quite low. The most frequently used yeast for the surface display of proteins is Saccharomyces cerevisiae. However, apart from its many advantages, Saccharomyces cerevisiae has some disadvantages, such as low robustness in industrial applications, hyperglycosylation of some heterologous proteins, and relatively low efficiency of surface display. Thus, in the recent years the display systems for alternative yeast hosts with better performances including Pichia pastoris, Hansenula polymorpha, Blastobotrys adeninivorans, Yarrowia lipolytica, Kluyveromyces marxianus, and others have been developed. Different strategies of surface display aimed to increase the amount of displayed protein, including new anchoring systems and new yeast hosts are reviewed in this paper.

Systematic Comparison of Cell Wall-Related Proteins of Different Yeasts

“…Therefore, it seems that rather complex biochemical pathways of GPI and β-1,6-glucan synthesis, proteins’ attachment to GPI, and eventually their translocation to β-1,6-glucan were not designed to attach particular protein(s) or sets of proteins, but rather to enable flexibility in adapting to different environments and conditions. In favor of this could also speak the fact that a rather simple protein modification, addition of the C-terminal GPI-anchoring signal, would result in binding of almost any homologous or heterologous protein in the cell wall through the GPI and β-1,6-glucan linkage [ 56 ]. Thus, addition of GPI-anchoring signals to different proteins could be an adaptation strategy in different environments.…”

Evolutionary Overview of Molecular Interactions and Enzymatic Activities in the Yeast Cell Walls

“…Besides, our comprehension of the variability of the cell wall composition and understanding of the molecular basis of cell wall integrity is crucial for the development of new biotechnological applications of fungi as well as the creation of new antifungal therapies. Examples of such applications are the use of antifungal drugs based on the inhibition of β-1,3-glucan synthesis [116,117], or expression of heterologous proteins directed to the cell wall and displayed at the cell surface [118], but further strategies applied in cell surface engineering could result in new biotechnology tools with various applications.…”

Molecular dynamics simulation and experimental study of the surface-display of SPA protein via Lpp-OmpA system for screening of IgG

“…Increasing the stability of SpA protein under harsh treatment/washing conditions is an important factor to increase the yield and quality of purified antibodies (Rigi et al 2019;Cherf and Cochran 2015). One of the major strategies to increase proteins stability is display of proteins on the surface of a live cell as an alternative to classic protein immobilization approach (Lozančić et al 2019;Grewal, et al 2016). The classical immobilization of recombinant proteins on the surface of a matrix (Khodaei et al 2018) is sometimes challenging because the protein may lose its conformation and consequently its function.…”

“…Increasing the stability of SpA protein under harsh treatment/washing conditions is an important factor to increase the yield and quality of purified antibodies (Rigi et al 2019 ; Cherf and Cochran 2015 ). One of the major strategies to increase proteins stability is display of proteins on the surface of a live cell as an alternative to classic protein immobilization approach (Lozančić et al 2019 ; Grewal, et al 2016 ).…”