Evaluation of Pyrophosphate‐Driven Proton Pumps in Saccharomyces cerevisiae under Stress Conditions
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/94d47e76-daa4-463f-8b8f-586d56878077
- author
- 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
- organization
- publishing date
- 2024-03-20
- 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}}, }