A carrier fusion significantly induces unfolded protein response in heterologous protein production by Aspergillus oryzae

2011 ◽  
Vol 92 (6) ◽  
pp. 1197-1206 ◽  
Author(s):  
Ayako Ohno ◽  
Jun-ichi Maruyama ◽  
Takashi Nemoto ◽  
Manabu Arioka ◽  
Katsuhiko Kitamoto
Keyword(s):  
Unfolded Protein Response ◽  
Aspergillus Oryzae ◽  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production ◽  
Unfolded Protein ◽  
Protein Response

The endoplasmic reticulum and unfolded protein response in the control of mammalian recombinant protein production

2014 ◽  
Vol 36 (8) ◽  
pp. 1581-1593 ◽  
Author(s):  
Hirra Hussain ◽  
Rodrigo Maldonado-Agurto ◽  
Alan J. Dickson
Keyword(s):  
Endoplasmic Reticulum ◽  
Recombinant Protein ◽  
Unfolded Protein Response ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Unfolded Protein ◽  
Protein Response

scholarly journals Improvement of Foreign-Protein Production in Aspergillus niger var. awamori by Constitutive Induction of the Unfolded-Protein Response

2003 ◽  
Vol 69 (12) ◽  
pp. 6979-6986 ◽  
Author(s):  
Mari Valkonen ◽  
Michael Ward ◽  
Huaming Wang ◽  
Merja Penttilä ◽  
Markku Saloheimo
Keyword(s):  
Aspergillus Niger ◽  
Unfolded Protein Response ◽  
Protein Production ◽  
Production Systems ◽  
Foreign Protein ◽  
Mrna Levels ◽  
Unfolded Protein ◽  
Novel Strategy ◽  
The Unfolded Protein Response ◽  
Protein Response

ABSTRACT Unfolded-protein response (UPR) denotes the upregulation of endoplasmic reticulum (ER)-resident chaperone and foldase genes and numerous other genes involved in secretory functions during the accumulation of unfolded proteins into the ER. Overexpression of individual foldases and chaperones has been used in attempts to improve protein production in different production systems. We describe here a novel strategy to improve foreign-protein production. We show that the constitutive induction of the UPR pathway in Aspergillus niger var. awamori can be achieved by expressing the activated form of the transcription factor hacA. This induction enhances the production of Trametes versicolor laccase by up to sevenfold and of bovine preprochymosin by up to 2.8-fold in this biotechnically important fungus. The regulatory range of UPR was studied by analyzing the mRNA levels of novel A. niger var. awamori genes involved in different secretory functions. This revealed both similarities and differences to corresponding studies in Saccharomyces cerevisiae.

scholarly journals Construction of Quintuple Protease and Double Amylase Gene Deletant for Heterologous Protein Production in Aspergillus oryzae KBN616

2015 ◽  
Vol 21 (3) ◽  
pp. 297-307 ◽  
Author(s):  
Noriyuki Kitamoto ◽  
Natsuko Ono ◽  
Shoko Yoshino-Yasuda
Keyword(s):  
Aspergillus Oryzae ◽  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production ◽  
Amylase Gene

scholarly journals CRISPR/Cas9 and Transgene Verification of Gene Involvement in Unfolded Protein Response and Recombinant Protein Production in Barley Grain

2021 ◽  
Vol 12 ◽  
Author(s):  
Michael Panting ◽  
Inger Baeksted Holme ◽  
Jón Már Björnsson ◽  
Yingxin Zhong ◽  
Henrik Brinch-Pedersen
Keyword(s):  
Recombinant Protein ◽  
Er Stress ◽  
Unfolded Protein Response ◽  
Recombinant Proteins ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Protein Accumulation ◽  
Unfolded Protein ◽  
Protein Response ◽  
Positive Effect

The use of plants as heterologous hosts to produce recombinant proteins has some intriguing advantages. There is, however, the potential of overloading the endoplasmic reticulum (ER) capacity when producing recombinant proteins in the seeds. This leads to an ER-stress condition and accumulating of unfolded proteins. The unfolded protein response (UPR) is activated to alleviate the ER-stress. With the aim to increase the yield of human epidermal growth factor (EGF) and mouse leukemia inhibitory factor (mLIF) in barley, we selected genes reported to have increased expression during ER-induced stress. The selected genes were calreticulin (CRT), protein disulfide isomerase (PDI), isopentenyl diphosphate isomerase (IPI), glutathione-s-transferase (GST), HSP70, HSP26, and HSP16.9. These were knocked out using CRISPR/Cas9 or overexpressed by conventional transgenesis. The generated homozygous barley lines were crossed with barley plants expressing EGF or mLIF and the offspring plants analyzed for EGF and mLIF protein accumulation in the mature grain. All manipulated genes had an impact on the expression of UPR genes when plantlets were subjected to tunicamycin (TN). The PDI knockout plant showed decreased protein body formation, with protein evenly distributed in the cells of the endosperm. The two genes, GST and IPI, were found to have a positive effect on recombinant protein production. mLIF expression was increased in a F2 homozygous GST knockout mutant background as compared to a F2 GST wild-type offspring. The overexpression of IPI in a F1 cross showed a significant increase in EGF expression. We demonstrate that manipulation of UPR related genes can have a positive effect on recombinant protein accumulation.

Functional and transcriptomic analysis of the key unfolded protein response transcription factor HacA in Aspergillus oryzae

Gene ◽  
2016 ◽  
Vol 593 (1) ◽  
pp. 143-153 ◽  
Author(s):  
Bin Zhou ◽  
Jingyi Xie ◽  
Xiaokai Liu ◽  
Bin Wang ◽  
Li Pan
Keyword(s):  
Transcription Factor ◽  
Unfolded Protein Response ◽  
Aspergillus Oryzae ◽  
Transcriptomic Analysis ◽  
Unfolded Protein ◽  
Protein Response

scholarly journals Effects of Inactivation and Constitutive Expression of the Unfolded- Protein Response Pathway on Protein Production in the Yeast Saccharomyces cerevisiae

2003 ◽  
Vol 69 (4) ◽  
pp. 2065-2072 ◽  
Author(s):  
Mari Valkonen ◽  
Merja Penttilä ◽  
Markku Saloheimo
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Unfolded Protein Response ◽  
Protein Production ◽  
Active Form ◽  
Constitutive Expression ◽  
Amylase Secretion ◽  
Unfolded Protein ◽  
Foreign Proteins ◽  
The Unfolded Protein Response ◽  
Protein Response

ABSTRACT One strategy to obtain better yields of secreted proteins has been overexpression of single endoplasmic reticulum-resident foldases or chaperones. We report here that manipulation of the unfolded-protein response (UPR) pathway regulator, HAC1, affects production of both native and foreign proteins in the yeast Saccharomyces cerevisiae. The effects of HAC1 deletion and overexpression on the production of a native protein, invertase, and two foreign proteins, Bacillus amyloliquefaciens α-amylase and Trichoderma reesei endoglucanase EGI, were studied. Disruption of HAC1 caused decreases in the secretion of both α-amylase (70 to 75% reduction) and EGI (40 to 50% reduction) compared to the secretion by the parental strain. Constitutive overexpression of HAC1 caused a 70% increase in α-amylase secretion but had no effect on EGI secretion. The invertase levels were twofold higher in the strain overexpressing HAC1. Also, the effect of the active form of T. reesei hac1 was tested in S. cerevisiae. hac1 expression caused a 2.4-fold increase in the secretion of α-amylase in S. cerevisiae and also slight increases in invertase and total protein production. Overexpression of both S. cerevisiae HAC1 and T. reesei hac1 caused an increase in the expression of the known UPR target gene KAR2 at early time points during cultivation.

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