Yeast synthetic biology for the production of recombinant therapeutic proteins

Abstract

The production of recombinant therapeutic proteins is one of the fast-growing areas of molecular medicine and currently plays an important role in treatment of several diseases. Yeasts are unicellular eukaryotic microbial host cells that offer unique advantages in producing biopharmaceutical proteins. Yeasts are capable of robust growth on simple media, readily accommodate genetic modifications, and incorporate typical eukaryotic post-translational modifications. Saccharomyces cerevisiae is a traditional baker's yeast that has been used as a major host for the production of biopharmaceuticals; however, several nonconventional yeast species including Hansenula polymorpha, Pichia pastoris, and Yarrowia lipolytica have gained increasing attention as alternative hosts for the industrial production of recombinant proteins. In this review, we address the established and emerging genetic tools and host strains suitable for recombinant protein production in various yeast expression systems, particularly focusing on current efforts toward synthetic biology approaches in developing yeast cell factories for the production of therapeutic recombinant proteins.

Genetic engineering approaches to improve posttranslational modification of biopharmaceuticals in different production platforms

“…Due to cell compartmentalization, yeast micro-organisms are capable of correct protein folding and further have the ability to perform typical eukaryotic PTMs including N-and, to some extent, O-linked glycosylation, phosphorylation, sulfation, and ubiquitination as described and reviewed by others (Irani, Kerkhoven, Shojaosadati, & Nielsen, 2015;Kim, Yoo, & Kang, 2015;Ptacek et al, 2005). Due to cell compartmentalization, yeast micro-organisms are capable of correct protein folding and further have the ability to perform typical eukaryotic PTMs including N-and, to some extent, O-linked glycosylation, phosphorylation, sulfation, and ubiquitination as described and reviewed by others (Irani, Kerkhoven, Shojaosadati, & Nielsen, 2015;Kim, Yoo, & Kang, 2015;Ptacek et al, 2005).…”

“…Nevertheless, high expression levels, less expensive and less complex bioprocessing, as well as well-characterized genetics with an extensive molecular toolbox including inducible promoters are the benefits of manufacturing in bacteria (Sahdev, Khattar, & Saini, 2008). glycosylation, phosphorylation, sulfation, and ubiquitination as described and reviewed by others (Irani, Kerkhoven, Shojaosadati, & Nielsen, 2015;Kim, Yoo, & Kang, 2015;Ptacek et al, 2005). Based on the lack of human-like glycosylation, yeast strains are mainly used for the commercial manufacturing of smaller proteins, hormones, and vaccines, with several products currently in the market (Cao et al, 2018;Mattanovich et al, 2011;Nielsen, 2013).…”

Genetic Manipulation of a Lipolytic Yeast Candida aaseri SH14 Using CRISPR-Cas9 System

“…Yeasts are unicellular eukaryotes with a long history of use in the biopharmaceutics and food industries [1,2]. Owing to their robust physiology in harsh conditions, such as elevated temperatures and low pH, many different yeasts have also been explored for the production of renewable bio-based chemicals to replace environmentally detrimental petrochemicals [3].…”

Transcriptional engineering of the glyceraldehyde‐3‐phosphate dehydrogenase promoter for improved heterologous protein production in Pichia pastoris

“…is a methylotrophic yeast that is widely used for heterologous protein production as it possesses many advantages such as having the ability to grow to very high cell densities on minimal media, expressing foreign proteins at very high levels either intracellularly and extracellularly, integration of expression plasmids at specific sites of the genome both in multiple or single copies and performing many posttranslational modifications (Çelik and Çalık, ). Due to its popularity, P. pastoris has been reviewed recently from several aspects such as protein folding and secretion (Damasceno et al, ; Puxbaum et al, ); fermentation strategies (Looser et al, ); genetic engineering approaches as well as synthetic biology tools (Kim et al, ; Li et al, ; Vogl et al, ; Vogl and Glieder, ; Wagner and Alper, ).…”