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Combined engineerin...
Combined engineering of disaccharide transport and phosphorolysis for enhanced ATP yield from sucrose fermentation in Saccharomyces cerevisiae
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Marques, W. L. (författare)
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Mans, R. (författare)
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Henderson, R. K. (författare)
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visa fler...
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Marella, E. R. (författare)
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Horst, J. T. (författare)
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Hulster, E. D. (författare)
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Poolman, B. (författare)
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Daran, J. -M (författare)
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Pronk, J. T. (författare)
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Gombert, A. K. (författare)
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- van Maris, Antonius J. A., Professor, 1976- (författare)
- KTH,Industriell bioteknologi
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(creator_code:org_t)
- Academic Press Inc. 2018
- 2018
- Engelska.
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Ingår i: Metabolic engineering. - : Academic Press Inc.. - 1096-7176 .- 1096-7184. ; 45, s. 121-133
- Relaterad länk:
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https://pure.rug.nl/...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Anaerobic industrial fermentation processes do not require aeration and intensive mixing and the accompanying cost savings are beneficial for production of chemicals and fuels. However, the free-energy conservation of fermentative pathways is often insufficient for the production and export of the desired compounds and/or for cellular growth and maintenance. To increase free-energy conservation during fermentation of the industrially relevant disaccharide sucrose by Saccharomyces cerevisiae, we first replaced the native yeast α-glucosidases by an intracellular sucrose phosphorylase from Leuconostoc mesenteroides (LmSPase). Subsequently, we replaced the native proton-coupled sucrose uptake system by a putative sucrose facilitator from Phaseolus vulgaris (PvSUF1). The resulting strains grew anaerobically on sucrose at specific growth rates of 0.09 ± 0.02 h−1 (LmSPase) and 0.06 ± 0.01 h−1 (PvSUF1, LmSPase). Overexpression of the yeast PGM2 gene, which encodes phosphoglucomutase, increased anaerobic growth rates on sucrose of these strains to 0.23 ± 0.01 h−1 and 0.08 ± 0.00 h−1, respectively. Determination of the biomass yield in anaerobic sucrose-limited chemostat cultures was used to assess the free-energy conservation of the engineered strains. Replacement of intracellular hydrolase with a phosphorylase increased the biomass yield on sucrose by 31%. Additional replacement of the native proton-coupled sucrose uptake system by PvSUF1 increased the anaerobic biomass yield by a further 8%, resulting in an overall increase of 41%. By experimentally demonstrating an energetic benefit of the combined engineering of disaccharide uptake and cleavage, this study represents a first step towards anaerobic production of compounds whose metabolic pathways currently do not conserve sufficient free-energy.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Bioenergi (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Bioenergy (hsv//eng)
Nyckelord
- ATP
- Chemostat
- Facilitated diffusion
- Free-energy conservation
- Phosphoglucomutase
- Yeast physiology
- Adenosinetriphosphate
- Biomass
- Chemostats
- Ecology
- Energy conservation
- Enzyme activity
- Fermentation
- Free energy
- Industrial chemicals
- Phosphorylation
- Yeast
- Facilitated diffusions
- Industrial fermentation
- Leuconostoc mesenteroides
- Phaseolus vulgaris
- Specific growth rate
- Sucrose phosphorylase
- Yeast physiologies
- Sugar (sucrose)
- adenosine triphosphate
- disaccharide
- glucose 6 phosphate
- hydrolase
- phosphorylase
- sucrose
- alkalinization
- anaerobic growth
- Arabidopsis thaliana
- Article
- carbon source
- cell suspension
- cellular distribution
- chemical reaction
- controlled study
- fungus growth
- genetic background
- metabolic engineering
- microscopy
- molecular biology
- nonhuman
- pea
- priority journal
- protein content
- proton motive force
- rice
- Saccharomyces cerevisiae
- sucrose metabolism
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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- Av författaren/redakt...
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Marques, W. L.
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Mans, R.
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Henderson, R. K.
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Marella, E. R.
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Horst, J. T.
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Hulster, E. D.
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visa fler...
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Poolman, B.
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Daran, J. -M
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Pronk, J. T.
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Gombert, A. K.
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van Maris, Anton ...
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visa färre...
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