ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production
Abstract Background Enforced ATP wasting has been recognized as a promising metabolic engineering strategy to enhance the microbial production of metabolites that are coupled to ATP generation. It also appears to be a suitable approach to improve production of ethanol by Saccharomyces cerevisiae. In...
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doaj-64f5503a85bb45e7947444d0d5d58dee2020-11-25T04:05:21ZengBMCBiotechnology for Biofuels1754-68342020-11-0113111210.1186/s13068-020-01822-9ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol productionAhmed Zahoor0Katrin Messerschmidt1Simon Boecker2Steffen Klamt3Analysis and Redesign of Biological Networks, Max Planck Institute for Dynamics of Complex Technical SystemsAnalysis and Redesign of Biological Networks, Max Planck Institute for Dynamics of Complex Technical SystemsAnalysis and Redesign of Biological Networks, Max Planck Institute for Dynamics of Complex Technical SystemsAnalysis and Redesign of Biological Networks, Max Planck Institute for Dynamics of Complex Technical SystemsAbstract Background Enforced ATP wasting has been recognized as a promising metabolic engineering strategy to enhance the microbial production of metabolites that are coupled to ATP generation. It also appears to be a suitable approach to improve production of ethanol by Saccharomyces cerevisiae. In the present study, we constructed different S. cerevisiae strains with heterologous expression of genes of the ATP-hydrolyzing F1-part of the ATPase enzyme to induce enforced ATP wasting and quantify the resulting effect on biomass and ethanol formation. Results In contrast to genomic integration, we found that episomal expression of the αβγ subunits of the F1-ATPase genes of Escherichia coli in S. cerevisiae resulted in significantly increased ATPase activity, while neither genomic integration nor episomal expression of the β subunit from Trichoderma reesei could enhance ATPase activity. When grown in minimal medium under anaerobic growth-coupled conditions, the strains expressing E. coli’s F1-ATPase genes showed significantly improved ethanol yield (increase of 10% compared to the control strain). However, elevated product formation reduces biomass formation and, therefore, volumetric productivity. We demonstrate that this negative effect can be overcome under growth-decoupled (nitrogen-starved) operation with high and constant biomass concentration. Under these conditions, which mimic the second (production) phase of a two-stage fermentation process, the ATPase-expressing strains showed significant improvement in volumetric productivity (up to 111%) compared to the control strain. Conclusions Our study shows that expression of genes of the F1-portion of E. coli’s ATPase induces ATPase activity in S. cerevisiae and can be a promising way to improve ethanol production. This ATP-wasting strategy can be easily applied to other metabolites of interest, whose formation is coupled to ATP generation.http://link.springer.com/article/10.1186/s13068-020-01822-9Enforced ATP wastingEthanolSaccharomyces cerevisiaeF1-ATPaseMetabolic engineering |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ahmed Zahoor Katrin Messerschmidt Simon Boecker Steffen Klamt |
spellingShingle |
Ahmed Zahoor Katrin Messerschmidt Simon Boecker Steffen Klamt ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production Biotechnology for Biofuels Enforced ATP wasting Ethanol Saccharomyces cerevisiae F1-ATPase Metabolic engineering |
author_facet |
Ahmed Zahoor Katrin Messerschmidt Simon Boecker Steffen Klamt |
author_sort |
Ahmed Zahoor |
title |
ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production |
title_short |
ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production |
title_full |
ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production |
title_fullStr |
ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production |
title_full_unstemmed |
ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production |
title_sort |
atpase-based implementation of enforced atp wasting in saccharomyces cerevisiae for improved ethanol production |
publisher |
BMC |
series |
Biotechnology for Biofuels |
issn |
1754-6834 |
publishDate |
2020-11-01 |
description |
Abstract Background Enforced ATP wasting has been recognized as a promising metabolic engineering strategy to enhance the microbial production of metabolites that are coupled to ATP generation. It also appears to be a suitable approach to improve production of ethanol by Saccharomyces cerevisiae. In the present study, we constructed different S. cerevisiae strains with heterologous expression of genes of the ATP-hydrolyzing F1-part of the ATPase enzyme to induce enforced ATP wasting and quantify the resulting effect on biomass and ethanol formation. Results In contrast to genomic integration, we found that episomal expression of the αβγ subunits of the F1-ATPase genes of Escherichia coli in S. cerevisiae resulted in significantly increased ATPase activity, while neither genomic integration nor episomal expression of the β subunit from Trichoderma reesei could enhance ATPase activity. When grown in minimal medium under anaerobic growth-coupled conditions, the strains expressing E. coli’s F1-ATPase genes showed significantly improved ethanol yield (increase of 10% compared to the control strain). However, elevated product formation reduces biomass formation and, therefore, volumetric productivity. We demonstrate that this negative effect can be overcome under growth-decoupled (nitrogen-starved) operation with high and constant biomass concentration. Under these conditions, which mimic the second (production) phase of a two-stage fermentation process, the ATPase-expressing strains showed significant improvement in volumetric productivity (up to 111%) compared to the control strain. Conclusions Our study shows that expression of genes of the F1-portion of E. coli’s ATPase induces ATPase activity in S. cerevisiae and can be a promising way to improve ethanol production. This ATP-wasting strategy can be easily applied to other metabolites of interest, whose formation is coupled to ATP generation. |
topic |
Enforced ATP wasting Ethanol Saccharomyces cerevisiae F1-ATPase Metabolic engineering |
url |
http://link.springer.com/article/10.1186/s13068-020-01822-9 |
work_keys_str_mv |
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