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

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    133 Citations (Scopus)

    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

    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

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