Enhanced production of antithrombotic hirudin by coexpression of Pdi1 and Ero1 in recombinant Saccharomyces cerevisiae

Kim, Myoung-Dong; Park, Eun-Hee; Cho, Jea-Won; Kim, Jin‐Chul; Cho, Seong-Min; Han, Miran; Seo, Jin‐Ho

doi:10.1016/j.jbiotec.2007.07.858

Cited by 4 publications

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although there is insufficient evidence regarding the effect of ERO overexpression on target proteins, some studies have shown to support the hypothesis that increased amount of ERO can increase the q p by maintaining oxidized PDI and boosting oxidative folding. In yeast, it has been shown that increased concentration of Ero1p enhances the rate of RNase A oxidative refolding (Tu et al, 2000), and co‐expression of Pdi1p and Ero1p increases the productivity of hirudin (Kim et al, 2007). In mammalian cells, Ero1‐L accelerated the oxidative folding of immunoglobulin in the ER (Mezghrani et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Effect of inducible co‐overexpression of protein disulfide isomerase and endoplasmic reticulum oxidoreductase on the specific antibody productivity of recombinant Chinese hamster ovary cells

Mohan

Lee

2010

Biotech & Bioengineering

View full text Add to dashboard Cite

To enhance specific antibody (Ab) productivity (q(Ab)) of recombinant Chinese hamster ovary (rCHO) cells, post-translational limitations in the endoplasmic reticulum during antibody production should be relieved. Previously, we reported that overexpression of protein disulfide isomerase (PDI), which catalyzes disulfide bond exchanges and assists in protein folding of newly synthesized proteins, enhanced q(Ab) of rCHO cells by about 27% (Mohan et al., 2007, Biotechnol Bioeng 98:611-615) . Since the rate limiting step in disulfide bond formation is found to be the regeneration of oxidized PDI, the oxidation state of PDI, as well as the amount of PDI, might be important. Endoplasmic reticulum oxidoreductase (ERO1L) maintains PDI in an oxidized state so that disulfide bond formation occurs. Here, PDI and its helper protein, ERO1L were overexpressed in rCHO cells producing an Ab in an attempt to ease the bottleneck in disulfide bond formation, and hence, Ab folding and secretion. Transient expression of ERO1L alone and with PDI resulted in enhanced q(Ab) by 37% and 55%, respectively. In contrast, under stable inducible co-overexpression of PDI and ERO1L, the q(Ab) was unaffected or negatively affected by varying degrees, depending on the individual expression levels of these genes. In stable clones with altered oxidation state of PDI due to co-overexpression of PDI and ERO1L, secretion of Ab was hindered and PDI-associated retention of Ab was seen in the cells. Under transient gene expression, secretion of Ab was not compromised. The data presented here suggests a possible mechanism of PDI/ERO1L interaction with the target Ab and shows how the expression levels of these proteins could affect the q(Ab) of this Ab-producing rCHO cell line.

show abstract

Section: Introductionmentioning

confidence: 99%

Effect of inducible co‐overexpression of protein disulfide isomerase and endoplasmic reticulum oxidoreductase on the specific antibody productivity of recombinant Chinese hamster ovary cells

Mohan

Lee

2010

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

Sulfhydryl oxidases: sources, properties, production and applications

Faccio

Nivala

Kruus

et al. 2011

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

The formation of disulfide bonds in proteins and small molecules can greatly affect their functionality. Sulfhydryl oxidases (SOXs) are enzymes capable of oxidising the free sulfhydryl groups in proteins and thiol-containing small molecules by using molecular oxygen as an electron acceptor. SOXs have been isolated from the intracellular compartments of many organisms, but also secreted SOXs are known. These latter enzymes are generally active on small compounds and their physiological role is unknown, whereas the intracellular enzymes prefer proteins as substrates and are involved in protein folding. An increasing number of scientific publications and patent applications on SOXs have been published in recent years. The present mini-review provides an up-to-date summary of SOXs from various families, their production and their actual or suggested applications. The sequence features and domain organisation of the characterised SOXs are reviewed, and special attention is paid to the physicochemical features of the enzymes. A review of patents and patent applications regarding this class of enzymes is also provided.

show abstract

Hirudin variants production by genetic engineered microbial factory

Zhang

Lan

2018

Biotechnology and Genetic Engineering Reviews

View full text Add to dashboard Cite

Hirudin was discovered as an active anticoagulant in leech extracts almost 60 years ago. Since their initial discovery, hirudin and its variants have been produced with various anti-thrombotic, cancer cell inhibition, diabetic cataract treatment and anti-fatigue activities. Some hirudin variants have been approved for clinical use and released into the marketplace. Recent progress has seen made in relation to hirudin variants expressed in several well-established microbial hosts, including Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and others, with high levels of activity and yield. This review summarizes the current progress on hirudin production using microbial producers, and considers the outlook for future development.

show abstract

Enhanced production of antithrombotic hirudin by coexpression of Pdi1 and Ero1 in recombinant Saccharomyces cerevisiae

Cited by 4 publications

References 0 publications

Effect of inducible co‐overexpression of protein disulfide isomerase and endoplasmic reticulum oxidoreductase on the specific antibody productivity of recombinant Chinese hamster ovary cells

Effect of inducible co‐overexpression of protein disulfide isomerase and endoplasmic reticulum oxidoreductase on the specific antibody productivity of recombinant Chinese hamster ovary cells

Sulfhydryl oxidases: sources, properties, production and applications

Hirudin variants production by genetic engineered microbial factory

Contact Info

Product

Resources

About