Overexpression of HAC1 gene increased levels of both intracellular and secreted human kringle fragment in Saccharomyces cerevisiae

Lee, Taehee; Bae, Yi-Hyun; Kim, Myoung-Dong; Seo, Jin‐Ho

doi:10.1016/j.procbio.2012.09.006

Cited by 8 publications

(4 citation statements)

References 34 publications

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“…Since ER stress is well characterized by overexpression of HAC1 , a transcription factor involved in unfolded protein response (UPR) and is considered as a marker for ER stress. 48 In our study, we have demonstrated that HAC1 expression is increased in SL treated cells. Carvacrol, an antifungal compound is known to exert its antifungal activity by disrupting calcium homeostasis and increasing HAC1 expression.…”

Section: Discussionsupporting

confidence: 53%

Sophorolipid exhibits antifungal activity by ROS mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways in Candida albicans

et al. 2019

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Section: Discussionsupporting

confidence: 53%

Sophorolipid exhibits antifungal activity by ROS mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways in Candida albicans

et al. 2019

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“…Another strategy has been to overexpress the main transcriptional regulator of the UPR, Hac1p, which has been successfully demonstrated in S. cerevisiae and Aspergillus niger var. awamori [ 46 , 47 ] . In P. pastoris overexpression of Hac1p in a single copy strain yielded mixed results depending on the heterologous protein in question [ 30 ].…”

Section: Introductionmentioning

confidence: 99%

Can too many copies spoil the broth?

Polizzi

2013

Microb Cell Fact

105

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The success of Pichia pastoris as a heterologous expression system lies predominantly in the impressive yields that can be achieved due to high volumetric productivity. However, low specific productivity still inhibits the potential success of this platform. Multi-(gene) copy clones are potentially a quick and convenient method to increase recombinant protein titer, yet they are not without their pitfalls. It has been more than twenty years since the first reported use of multi-copy clones and it is still an active area of research to find the fastest and most efficient method for generating these strains. It has also become apparent that there is not always a linear correlation between copy number and protein titer, leading to in-depth investigations into how to minimize the negative impact of secretory stress and achieve clonal stability.

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“…Lee et al studied the effects of overexpression of a UPR pathway gene in recombinant S. cerevisiae strains harbouring 16 copies of the gene coding for human kringle protein LK8. Their results showed that Hac1p promotes LK8 protein production as well as cell growth in recombinant S. cerevisiae 28 . In the current study, HAC1 was overexpressed in recombinant S. cerevisiae strains containing one, eight (the highest enzyme activity), or 22 (the highest number of copies) copies of xynB , with overexpression of HAC1 in the wild-type strain used as a control.…”

Section: Discussionmentioning

confidence: 99%

Expression and function of an Hac1-regulated multi-copy xylanase gene in Saccharomyces cerevisiae

Bao

Chen

et al. 2020

Sci Rep

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Saccharomyces cerevisiae -based expression systems, which rely on safe, food-grade strains, are low cost, simple to operate, and can be used for large-scale fermentation. However, low levels of foreign protein expression by S. cerevisiae have limited their widespread application. The ability of the endoplasmic reticulum (ER) to fold and process foreign proteins is an important factor restricting the expression of foreign proteins. In the current study, the effects of transcription factor Hac1p, which is involved in the unfolded protein response pathway, on S. cerevisiae -based expression of xylanase gene xynB from Aspergillus niger were examined. Overlap extension polymerase chain reaction (PCR), rDNA integration and droplet digital PCR technology were used to generate a S. cerevisiae strain (S8) containing eight copies of xynB , allowing high-yield secretory expression of xylanase. The effects of subsequent overexpression of HAC1 in strain S8 on the expression of genes associated with protein folding in the ER were then examined using the GeXP system. Results confirmed the constitutive secretory expression of the multiple copies of xynB following rDNA-based integration of the expression cassette, with a maximum xylanase yield of 325 U/mL. However, overexpression of HAC1 further improved xylanase production by strain S8, resulting in a yield of 381 U/mL.

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Overexpression of HAC1 gene increased levels of both intracellular and secreted human kringle fragment in Saccharomyces cerevisiae

Cited by 8 publications

References 34 publications

Sophorolipid exhibits antifungal activity by ROS mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways in Candida albicans

Sophorolipid exhibits antifungal activity by ROS mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways in Candida albicans

Can too many copies spoil the broth?

Expression and function of an Hac1-regulated multi-copy xylanase gene in Saccharomyces cerevisiae

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