Yeast synthetic biology for designed cell factories producing secretory recombinant proteins

Abstract

Yeasts are prominent hosts for the production of recombinant proteins from industrial enzymes to therapeutic proteins. Particularly, the similarity of protein secretion pathways between these unicellular eukaryotic microorganisms and higher eukaryotic organisms has made them a preferential host to produce secretory recombinant proteins. However, there are several bottlenecks, in terms of quality and quantity, restricting their use as secretory recombinant protein production hosts. In this mini-review, we discuss recent developments in synthetic biology approaches to constructing yeast cell factories endowed with enhanced capacities of protein folding and secretion as well as designed targeted post-translational modification process functions. We focus on the new genetic tools for optimizing secretory protein expression, such as codon-optimized synthetic genes, combinatory synthetic signal peptides and copy number-controllable integration systems, and the advanced cellular engineering strategies, including endoplasmic reticulum and protein trafficking pathway engineering, synthetic glycosylation, and cell wall engineering, for improving the quality and yield of secretory recombinant proteins.

Modulation of heterologous protein secretion in the thermotolerant methylotrophic yeast Ogataea thermomethanolica TBRC 656 by CRISPR-Cas9 system

“…Like other yeasts, efficient secretion is a major bottleneck for heterologous protein production, in which heterologous proteins are often poorly secreted, leading to intracellular accumulation, which triggers endoplasmic reticulum (ER) stress [ 8 , 9 ]. Therefore, improving the secretory capacity through the engineering of secretory pathways could improve protein production, for example, by the fusion of signal peptides to the heterologous protein, by the engineering of protein folding, and by the ER quality control and engineering of vacuolar sorting pathways [ 10 , 11 ].…”

Comparison of the Unfolded Protein Response in Cellobiose Utilization of Recombinant Angel- and W303-1A-Derived Yeast Expressing β-Glucosidase

“…Saccharomyces cerevisiae is one of the most widely used cell factories and is utilized in biotechnological processes including the production of heterologous proteins, biofuels, and chemicals of medical or industrial interest ( Jouhten et al, 2016 ; Lian et al, 2018 ; Thak et al, 2020 ). In synthetic biology and metabolic engineering applications, yeast not only faces many kinds of stress, such as osmotic, heat, inhibitor, and nutrient starvation stress, but also endures a metabolic burden resulting from the heterologous production of enzymes to broaden the yeast’s substrate or product range ( Mattanovich et al, 2004 ; Wu et al, 2016; Brandt et al, 2021 ).…”

Development of synthetic biology tools to engineer Pichia pastoris as a chassis for the production of natural products

“…For example, the expression level of the target gene can account for more than 30% of the total proteins of P. pastoris , which is much higher than that in S. cerevisiae [ 8 ]. In addition, hyperglycosylation is another concern for expressing eukaryotic proteins in the S. cerevisiae expression system [ 9 ]. Natural product biosynthetic pathway enzymes (i.e.…”