Recombinant Protein Production in Yeasts

Abstract

Recombinant protein production is a multibillion-dollar market. The development of a new product begins with the choice of a production host. While one single perfect host for every protein does not exist, several expression systems ranging from bacterial hosts to mammalian cells have been established. Among them, yeast cell factories combine the advantages of being single cells, such as fast growth and easy genetic manipulation, as well as eukaryotic features including a secretory pathway leading to correct protein processing and post-translational modifications. In this respect, especially the engineering of yeast glycosylation to produce glycoproteins of human-like glycan structures is of great interest. Additionally, different attempts of cellular engineering as well as the design of different production processes that are leading to improved productivities are presented. With the advent of cheaper next-generation sequencing techniques, systems biotechnology approaches focusing on genome scale analyses will advance and accelerate yeast cell factories and thus recombinant protein production processes in the near future. In this review we summarize advantages and limitations of the main and most promising yeast hosts, including Saccharomyces cerevisiae, Pichia pastoris, and Hansenula polymorpha as those presently used in large scale production of heterologous proteins.

Multistep processing of the secretion leader of the extracellular protein Epx1 in Pichia pastoris and implications for protein localization

“…Yeasts are commonly used host organisms for production of a wide range of heterologous proteins, such as enzymes, vaccines, hormones and biopharmaceutical proteins. Most of these proteins are secreted in their native environment; thus, should preferably also be secreted by the recombinant expression host (Macauley-Patrick et al, 2005;Mattanovich et al, 2012). The methylotrophic yeast Pichia pastoris (synonym Komagataella sp.…”

“…By far the most-used and best-studied secretion leader in yeast is the MFa pre-pro-leader originating from S. cerevisiae, which has been used for production of secretory proteins with and without an EAEA (Glu-Ala) 2 overhang in various yeast species (Daly & Hearn, 2005;Ghosalkar et al, 2008;Mattanovich et al, 2012). Unfortunately, both variants of MFa suffer from inefficient processing by Kex2 and/or Ste13, which leaves unwanted overhangs at the N-termini of the produced recombinant proteins and leads to heterogeneous products (among others: Almeida et al, 2001;Hashimoto et al, 1998;Steinlein et al, 1995;Zhao et al, 2009).…”

Overview

“…In the early 1980s this was the only yeast species with significant molecular genetic characterization which explains its wide commercial use in the following years for production of human insulin and hepatitis B surface antigen. It has turned out however that other yeasts are more efficient in the production of many recombinant proteins [31][32][33]. A current literature survey indicates that most work on recombinant protein production in yeasts is performed with P. pastoris and H. polymorpha, followed by S. cerevisiae and Y. lipolytica.…”

Structural Biology of Glycoproteins

“…Yeast have been used for the production of human glycoproteins, with the most popular expression hosts being Saccharomyces cerevisiae and Pichia pastoris [24][25]. Recombinant proteins are secreted and post-translationally glycosylated to give glycoforms that are sensitive to endo H or endo F1 treatment and are therefore suitable for crystallization ( Figure 4B).…”