Effects of inactivation and constitutive expression of the unfolded-protein response pathway on protein production in the yeast Saccharomyces cerevisiae

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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.

Original languageEnglish
Pages (from-to)2065-2072
Number of pages8
JournalApplied and Environmental Microbiology
Volume69
Issue number4
DOIs
Publication statusPublished - 1 Apr 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

unfolded protein response
Unfolded Protein Response
Amylases
beta-Fructofuranosidase
yeast
Saccharomyces cerevisiae
inactivation
amylases
Trichoderma reesei
Yeasts
secretion
yeasts
beta-fructofuranosidase
protein
Proteins
proteins
Trichoderma
Fungal Proteins
Cellulase
Bacillus amyloliquefaciens

Keywords

  • Proteins
  • strain
  • Bacillus amyloliquefaciens
  • Unfolded-protein response (UPR)
  • amylase
  • beta fructofuranosidase
  • gene product
  • glucan synthase
  • protein hac1
  • yeast
  • fungi
  • Hypocrea jecorina
  • Saccharomyces
  • esSaccharomyces cerevisiae
  • Trichoderma

Cite this

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title = "Effects of inactivation and constitutive expression of the unfolded-protein response pathway on protein production in the yeast Saccharomyces cerevisiae",
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.",
keywords = "Proteins, strain, Bacillus amyloliquefaciens, Unfolded-protein response (UPR), amylase, beta fructofuranosidase, gene product, glucan synthase, protein hac1, yeast, fungi, Hypocrea jecorina, Saccharomyces, esSaccharomyces cerevisiae, Trichoderma",
author = "Mari Valkonen and Merja Penttil{\"a} and Markku Saloheimo",
year = "2003",
month = "4",
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journal = "Applied and Environmental Microbiology",
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TY - JOUR

T1 - Effects of inactivation and constitutive expression of the unfolded-protein response pathway on protein production in the yeast Saccharomyces cerevisiae

AU - Valkonen, Mari

AU - Penttilä, Merja

AU - Saloheimo, Markku

PY - 2003/4/1

Y1 - 2003/4/1

N2 - 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.

AB - 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.

KW - Proteins

KW - strain

KW - Bacillus amyloliquefaciens

KW - Unfolded-protein response (UPR)

KW - amylase

KW - beta fructofuranosidase

KW - gene product

KW - glucan synthase

KW - protein hac1

KW - yeast

KW - fungi

KW - Hypocrea jecorina

KW - Saccharomyces

KW - esSaccharomyces cerevisiae

KW - Trichoderma

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U2 - 10.1128/AEM.69.4.2065-2072.2003

DO - 10.1128/AEM.69.4.2065-2072.2003

M3 - Article

VL - 69

SP - 2065

EP - 2072

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 4

ER -