Modifications to the ADH1 promoter of Saccharomyces cerevisiae for efficient production of heterologous proteins

Laura Ruohonen (Corresponding Author), Markku Aalto, Sirkka Keränen

Research output: Contribution to journalArticleScientificpeer-review

54 Citations (Scopus)

Abstract

The promoter of alcohol dehydrogenase I of the yeast Saccharomyces cerevisiae was studied using Bacillus amyloliquefaciens α-amylase as a marker protein. On glucose, activity of the original ADH1 promoter decreases during late exponential, ethanol production growth phase. When 1100 bp (from −414 bp to −1500 bp) of the upstream sequence are deleted, activity increases into the late ethanol consumption phase but the promoter becomes active only after ethanol production growth phase (Ruohonen et al. (1991) Yeast 7, 337–346). We have now restored 300 bp (from − 414 bp to − 700 bp) upstream of the deletion site and obtained expression from the ADH1 promoter throughout the yeast growth cycle. The restored sequence allowed a-amylase expression to start during early exponential growth phase indicating that it is required for activation of the ADH1 promoter during ethanol production growth phase, possibly through glucose induction. On ethanol, all the promoters were active, but the short promoter was temporally activated first, suggesting that the restored sequence is not required for promoter activity during early oxidative growth.
Original languageEnglish
Pages (from-to)193-203
JournalJournal of Biotechnology
Volume39
Issue number3
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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Yeast
Saccharomyces cerevisiae
Proteins
Ethanol
Growth
Amylases
Yeasts
Glucose
Alcohol Dehydrogenase
Bacilli
Alcohols
Chemical activation

Cite this

Ruohonen, Laura ; Aalto, Markku ; Keränen, Sirkka. / Modifications to the ADH1 promoter of Saccharomyces cerevisiae for efficient production of heterologous proteins. In: Journal of Biotechnology. 1995 ; Vol. 39, No. 3. pp. 193-203.
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abstract = "The promoter of alcohol dehydrogenase I of the yeast Saccharomyces cerevisiae was studied using Bacillus amyloliquefaciens α-amylase as a marker protein. On glucose, activity of the original ADH1 promoter decreases during late exponential, ethanol production growth phase. When 1100 bp (from −414 bp to −1500 bp) of the upstream sequence are deleted, activity increases into the late ethanol consumption phase but the promoter becomes active only after ethanol production growth phase (Ruohonen et al. (1991) Yeast 7, 337–346). We have now restored 300 bp (from − 414 bp to − 700 bp) upstream of the deletion site and obtained expression from the ADH1 promoter throughout the yeast growth cycle. The restored sequence allowed a-amylase expression to start during early exponential growth phase indicating that it is required for activation of the ADH1 promoter during ethanol production growth phase, possibly through glucose induction. On ethanol, all the promoters were active, but the short promoter was temporally activated first, suggesting that the restored sequence is not required for promoter activity during early oxidative growth.",
author = "Laura Ruohonen and Markku Aalto and Sirkka Ker{\"a}nen",
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Modifications to the ADH1 promoter of Saccharomyces cerevisiae for efficient production of heterologous proteins. / Ruohonen, Laura (Corresponding Author); Aalto, Markku; Keränen, Sirkka.

In: Journal of Biotechnology, Vol. 39, No. 3, 1995, p. 193-203.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Modifications to the ADH1 promoter of Saccharomyces cerevisiae for efficient production of heterologous proteins

AU - Ruohonen, Laura

AU - Aalto, Markku

AU - Keränen, Sirkka

N1 - Project code: BEL4301

PY - 1995

Y1 - 1995

N2 - The promoter of alcohol dehydrogenase I of the yeast Saccharomyces cerevisiae was studied using Bacillus amyloliquefaciens α-amylase as a marker protein. On glucose, activity of the original ADH1 promoter decreases during late exponential, ethanol production growth phase. When 1100 bp (from −414 bp to −1500 bp) of the upstream sequence are deleted, activity increases into the late ethanol consumption phase but the promoter becomes active only after ethanol production growth phase (Ruohonen et al. (1991) Yeast 7, 337–346). We have now restored 300 bp (from − 414 bp to − 700 bp) upstream of the deletion site and obtained expression from the ADH1 promoter throughout the yeast growth cycle. The restored sequence allowed a-amylase expression to start during early exponential growth phase indicating that it is required for activation of the ADH1 promoter during ethanol production growth phase, possibly through glucose induction. On ethanol, all the promoters were active, but the short promoter was temporally activated first, suggesting that the restored sequence is not required for promoter activity during early oxidative growth.

AB - The promoter of alcohol dehydrogenase I of the yeast Saccharomyces cerevisiae was studied using Bacillus amyloliquefaciens α-amylase as a marker protein. On glucose, activity of the original ADH1 promoter decreases during late exponential, ethanol production growth phase. When 1100 bp (from −414 bp to −1500 bp) of the upstream sequence are deleted, activity increases into the late ethanol consumption phase but the promoter becomes active only after ethanol production growth phase (Ruohonen et al. (1991) Yeast 7, 337–346). We have now restored 300 bp (from − 414 bp to − 700 bp) upstream of the deletion site and obtained expression from the ADH1 promoter throughout the yeast growth cycle. The restored sequence allowed a-amylase expression to start during early exponential growth phase indicating that it is required for activation of the ADH1 promoter during ethanol production growth phase, possibly through glucose induction. On ethanol, all the promoters were active, but the short promoter was temporally activated first, suggesting that the restored sequence is not required for promoter activity during early oxidative growth.

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