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Evaluation of Pyrophosphate‐Driven Proton Pumps in Saccharomyces cerevisiae under Stress Conditions

Sreenivas, Krishnan LU ; Eisentraut, Leon ; Brink, Daniel P. LU ; Persson, Viktor C. LU orcid ; Carlquist, Magnus LU ; Gorwa-Grauslund, Marie F. LU and van Niel, Ed W. J. LU orcid (2024) In Microorganisms 12(3).
Abstract
In Saccharomyces cerevisiae, pH homeostasis is reliant on ATP due to the use of proton-translocating ATPase (H+-ATPase) which constitutes a major drain within cellular ATP supply. Here, an exogenous proton-translocating pyrophosphatase (H+-PPase) from Arabidopsis thaliana, which uses inorganic pyrophosphate (PPi) rather than ATP, was evaluated for its effect on reducing the ATP burden. The H+-Ppase was localized to the vacuolar membrane or to the cell membrane, and their impact was studied under acetate stress at a low pH. Biosensors (pHluorin and mQueen-2m) were used to observe changes in intracellular pH (pHi) and ATP levels during growth on either glucose or xylose. A significant... (More)
In Saccharomyces cerevisiae, pH homeostasis is reliant on ATP due to the use of proton-translocating ATPase (H+-ATPase) which constitutes a major drain within cellular ATP supply. Here, an exogenous proton-translocating pyrophosphatase (H+-PPase) from Arabidopsis thaliana, which uses inorganic pyrophosphate (PPi) rather than ATP, was evaluated for its effect on reducing the ATP burden. The H+-Ppase was localized to the vacuolar membrane or to the cell membrane, and their impact was studied under acetate stress at a low pH. Biosensors (pHluorin and mQueen-2m) were used to observe changes in intracellular pH (pHi) and ATP levels during growth on either glucose or xylose. A significant improvement of 35% in the growth rate at a pH of 3.7 and 6 g·L−1 acetic acid stress was observed in the vacuolar membrane H+-PPase strain compared to the parent strain. ATP levels were elevated in the same strain during anaerobic glucose and xylose fermentations. During anaerobic xylose fermentations, co-expression of pHluorin and a vacuolar membrane H+-PPase improved the growth characteristics by means of an improved growth rate (11.4%) and elongated logarithmic growth duration. Our study identified a potential method for improving productivity in the use of S. cerevisiae as a cell factory under the harsh conditions present in industry. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Saccharomyces cerevisiae, pH homeostasis, ATP, proton-translocating ATPase, proton-translocating pyrophosphatase, pHluorin, mQueen-2m, acetic acid, xylose
in
Microorganisms
volume
12
issue
3
article number
625
pages
23 pages
publisher
MDPI AG
external identifiers
  • scopus:85189105336
  • pmid:38543676
ISSN
2076-2607
DOI
10.3390/microorganisms12030625
project
Molecular understanding of microbial growth constraints by the interconnection between the redox- and energy carrier formation fluxes
Improved redox- and energy metabolism of Saccharomyces cerevisiae on xylose: verifying an effective microbial cell factory
language
English
LU publication?
yes
id
94d47e76-daa4-463f-8b8f-586d56878077
date added to LUP
2024-03-20 19:46:01
date last changed
2025-06-12 07:18:58
@article{94d47e76-daa4-463f-8b8f-586d56878077,
  abstract     = {{In Saccharomyces cerevisiae, pH homeostasis is reliant on ATP due to the use of proton-translocating ATPase (H<sup>+</sup>-ATPase) which constitutes a major drain within cellular ATP supply. Here, an exogenous proton-translocating pyrophosphatase (H<sup>+</sup>-PPase) from Arabidopsis thaliana, which uses inorganic pyrophosphate (PP<sub>i</sub>) rather than ATP, was evaluated for its effect on reducing the ATP burden. The H<sup>+</sup>-Ppase was localized to the vacuolar membrane or to the cell membrane, and their impact was studied under acetate stress at a low pH. Biosensors (pHluorin and mQueen-2m) were used to observe changes in intracellular pH (pH<sub>i</sub>) and ATP levels during growth on either glucose or xylose. A significant improvement of 35% in the growth rate at a pH of 3.7 and 6 g·L<sup>−1</sup> acetic acid stress was observed in the vacuolar membrane H<sup>+</sup>-PPase strain compared to the parent strain. ATP levels were elevated in the same strain during anaerobic glucose and xylose fermentations. During anaerobic xylose fermentations, co-expression of pHluorin and a vacuolar membrane H<sup>+</sup>-PPase improved the growth characteristics by means of an improved growth rate (11.4%) and elongated logarithmic growth duration. Our study identified a potential method for improving productivity in the use of S. cerevisiae as a cell factory under the harsh conditions present in industry.}},
  author       = {{Sreenivas, Krishnan and Eisentraut, Leon and Brink, Daniel P. and Persson, Viktor C. and Carlquist, Magnus and Gorwa-Grauslund, Marie F. and van Niel, Ed W. J.}},
  issn         = {{2076-2607}},
  keywords     = {{Saccharomyces cerevisiae; pH homeostasis; ATP; proton-translocating ATPase; proton-translocating pyrophosphatase; pHluorin; mQueen-2m; acetic acid; xylose}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{3}},
  publisher    = {{MDPI AG}},
  series       = {{Microorganisms}},
  title        = {{Evaluation of Pyrophosphate‐Driven Proton Pumps in <i>Saccharomyces cerevisiae </i>under Stress Conditions}},
  url          = {{https://lup.lub.lu.se/search/files/177716503/Sreenivas_et_al_2024_microorganisms-12-00625.pdf}},
  doi          = {{10.3390/microorganisms12030625}},
  volume       = {{12}},
  year         = {{2024}},
}