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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 ; Carlquist, Magnus LU ; Gorwa-Grauslund, Marie F. LU and van Niel, Ed W. J. LU (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
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
2024-04-16 15:37:05
@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}},
}