Bioreactor-Scale Strategies for the Production of Recombinant Protein in the Yeast Yarrowia lipolytica

Abstract

Recombinant protein production represents a multibillion-dollar market. Therefore, it constitutes an important research field both in academia and industry. The use of yeast as a cell factory presents several advantages such as ease of genetic manipulation, growth at high cell density, and the possibility of post-translational modifications. Yarrowia lipolytica is considered as one of the most attractive hosts due to its ability to metabolize raw substrate, to express genes at a high level, and to secrete protein in large amounts. In recent years, several reviews have been dedicated to genetic tools developed for this purpose. Though the construction of efficient cell factories for recombinant protein synthesis is important, the development of an efficient process for recombinant protein production in a bioreactor constitutes an equally vital aspect. Indeed, a sports car cannot drive fast on a gravel road. The aim of this review is to provide a comprehensive snapshot of process tools to consider for recombinant protein production in bioreactor using Y. lipolytica as a cell factory, in order to facilitate the decision-making for future strain and process engineering.

Comprehensive comparison of Yarrowia lipolytica and Pichia pastoris for production of Candida antarctica lipase B

“…These peculiar metabolic traits have been exploited to develop molecular tools for rProt synthesis and secretion 7 . When combined with efficient bioreactor process strategies, these tools have been successfully used for the production of a large number of rProt 8,9 . We have recently developed a novel set of expression vectors based on the promoter of EYK1 gene encoding erythrulose kinase 10,11 .…”

Impact of oxygen availability on heterologous geneexpression and polypeptide secretion dynamics in Yarrowia lipolytica‐based protein production platforms

“…In contrast, Y. lipolytica naturally secretes high levels of various native enzymes and hence its secretory pathway is naturally less prone to induce endoplasmic reticulum (ER) associated degradation (Theron, Vandermies, Telek, Steels, & Fickers, 2020). Although Y. lipolytica's native potential in this regard is highly competitive (alkaline extracellular protease can be secreted up to 2 g L −1 ; Le Dall, Nicaud, & Gaillardin, 1994;Ogrydziak, 2013), it is interesting to systematically study different parameters that enable further enhancement of this trait by both bioprocessing (Vandermies & Fickers, 2019) and genetic engineering approaches (Celi nska, Borkowska, Białas, Korpys, & Nicaud, 2018). While the impact of OA on physiology of Y. lipolytica has been already studied, the influence of OA on rProt synthesis and secretion has been largely neglected to date.…”

Application of combinatorial optimization strategies in synthetic biology

“…Chemical synthesis of protein molecules is prohibitively expensive. Several expression systems ranging from bacterial hosts to mammalian cells have been established 24 . However, only 25 of them have been produced at a bioreactor scale 24 .…”

“…Moreover, tweaking multiple factors can typically be critical to obtain an optimal output in a biological system 13 . Those may include overall structural state of chromatin and its domains 14 , the strength of transcriptional regulators controlling gene expression 8,15 , transcriptional terminators [16][17][18] , ribosome binding sites (RBS), biochemical properties of the protein(s) encoded by the recombinant genes [19][20][21] , the availability of cofactors for the correct functionality of enyzmes 22,23 , the genetic background of the host [24][25][26] , and the expression system itself (plasmid-based vs. chromosomal integration) 27 .…”