Established and Upcoming Yeast Expression Systems

Ergün, Burcu Gündüz; Hüccetoğulları, Damla; Öztürk, Sibel; Çelik, Eda; Çalı́k, Pınar

doi:10.1007/978-1-4939-9024-5_1

Cited by 30 publications

(16 citation statements)

References 513 publications

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“…S. pombe has considerable advantages as a host for the industrial-scale production of eukaryotic proteins related to transactions conserved in higher eukarya (RNA splicing, RNA interference pathway, etc.) and advantages in glycosylation of proteins (Celik and Calik 2012;Gunduz Ergun et al 2019). The pTIN system will allow convenient usage of a regulatable expression vector, where fermentation scale cultures may be grown in noninducing conditions and gene expression turned on by the addition of thiamine.…”

Section: Discussionmentioning

confidence: 99%

A lncRNA-regulated gene expression system with rapid induction kinetics in the fission yeast Schizosaccharomyces pombe

Garg

2020

RNA

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The fission yeast Schizosaccharomyces pombe is an excellent model organism for the study of eukaryotic cellular physiology. The organism is genetically tractable and several tools to study the functions of individual genes are available. One such tool is regulatable gene expression and overproduction of proteins. Limitations of currently available overexpression systems include delay in expression after induction, narrow dynamic range, and system-wide changes due to induction conditions. Here I describe a new long noncoding RNA (lncRNA)-regulated, thiamine-inducible expression system that integrates lncRNA-based transcriptional interference at the fission yeast tgp1 promoter with the fast repression kinetics of the thiamine-repressible nmt1 promoter. This hybrid system has rapid induction kinetics, broad dynamic range, and tunable expression via thiamine concentration. The lncRNA-regulated thiamine-inducible system will be advantageous for the study of individual genes and for potential applications in the production of heterologous proteins in fission yeast.

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Section: Discussionmentioning

confidence: 99%

A lncRNA-regulated gene expression system with rapid induction kinetics in the fission yeast Schizosaccharomyces pombe

Garg

2020

RNA

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“…Methylotrophic yeasts offer a repertoire of regulated, strong promoters that are naturally regulating MUT pathway genes (Ergün et al . 2019b ). Typically, the promoters of MUT pathway genes are tightly repressed on repressing carbon sources such as glucose and strongly induced when shifted to methanol.…”

Section: Gene Regulation and Tfsmentioning

confidence: 99%

What makes Komagataella phaffii non-conventional?

Ata

Ergün

Fickers

et al. 2021

FEMS Yeast Research

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The important industrial protein production host Komagataella phaffii (syn Pichia pastoris) is classified as a non-conventional yeast. But what exactly makes K. phaffii non-conventional? In this review we set out to address the main differences to the ‘conventional’ yeast Saccharomyces cerevisiae, but also pinpoint differences to other non-conventional yeasts used in biotechnology. Apart from its methylotrophic lifestyle, K. phaffii is a Crabtree-negative yeast species. But even within the methylotrophs, K. phaffii possesses distinct regulatory features such as glycerol-repression of the methanol-utilization pathway or the lack of nitrate assimilation. Rewiring of the transcriptional networks regulating carbon (and nitrogen) source utilization clearly contributes to our understanding of genetic events occurring during evolution of yeast species. The mechanisms of mating-type switching and the triggers of morphogenic phenotypes represent further examples for how K. phaffii is distinguished from the model yeast S. cerevisiae. With respect to heterologous protein production, K. phaffii features high secretory capacity but secretes only low amounts of endogenous proteins. Different to S. cerevisiae, the Golgi apparatus of K. phaffii is stacked like in mammals. While it is tempting to speculate that Golgi architecture is correlated to the high secretion levels or the different N-glycan structures observed in K. phaffii, there is recent evidence against this. We conclude that K. phaffii is a yeast with unique features that has a lot of potential to explore both fundamental research questions and industrial applications.

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“…Gene modification has been applied to construct a humanized N-linked glycosylation pathway in K. lactis . Besides the above conventional yeast species, other established yeast expression systems include Hansenula polymorpha (Gündüz Ergün et al, 2019 ; Fukunaga et al, 2021a , b ; Giorgetti et al, 2021 ; Gallo et al, 2022 ), Yarrowia lipolytica (Hamilton et al, 2013 ; Karbalaei et al, 2020 ; Jin et al, 2021 ), Schizosaccharomyces pombe (Kim et al, 2006 ; Katla et al, 2019 ; Lawton et al, 2021 ), Kluyveromyces marxianus (Lee et al, 2020 ), Candida albicans (Li et al, 2019 ), and Zygosaccharomyces cidri (Liu et al, 2009 ). The strategies to produce humanized glycosylated end-products were explored by using these yeast species.…”

Section: Introductionmentioning

confidence: 99%

Humanization of Yeasts for Glycan-Type End-Products

Li¹,

Shen²,

Chen³

et al. 2022

Front. Microbiol.

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Yeasts are often considered microorganisms for producing human therapeutic glycosylated end-products at an industrial scale. However, the products with non-humanized glycans limited their usage. Therefore, various methods to develop humanized glycosylated end-products have been widely reported in yeasts. To make full use of these methods, it is necessary to summarize the present research to find effective approaches to producing humanized products. The present research focuses on yeast species selection, glycosyltransferase deletion, expression of endoglycosidase, and expression of proteins with galactosylated and or sialylated glycans. Nevertheless, the yeasts will have growth defects with low bioactivity when the key enzymes are deleted. It is necessary to express the corresponding repairing protein. Compared with N-glycosylation, the function of yeast protein O-glycosylation is not well-understood. Yeast proteins have a wide variety of O-glycans in different species, and it is difficult to predict glycosylation sites, which limits the humanization of O-glycosylated yeast proteins. The future challenges include the following points: there are still many important potential yeasts that have never been tried to produce glycosylated therapeutic products. Their glycosylation pathway and related mechanisms for producing humanized glycosylated proteins have rarely been reported. On the other hand, the amounts of key enzymes on glycan pathways in human beings are significantly more than those in yeasts. Therefore, there is still a challenge to produce a large body of humanized therapeutic end-products in suitable yeast species, especially the protein with complex glycans. CRISPR-Cas9 system may provide a potential approach to address the important issue.

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Established and Upcoming Yeast Expression Systems

Cited by 30 publications

References 513 publications

A lncRNA-regulated gene expression system with rapid induction kinetics in the fission yeast Schizosaccharomyces pombe

A lncRNA-regulated gene expression system with rapid induction kinetics in the fission yeast Schizosaccharomyces pombe

What makes Komagataella phaffii non-conventional?

Humanization of Yeasts for Glycan-Type End-Products

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