Phosphate starvation is often applied as a tool to limit cell growth in
microbial production processes without hampering carbon and/or nitrogen
supply alternatively. This contribution focuses on the interplay of
process induced phosphate starvation and microbial performance studying
an l-tryptophan overproducing Escherichia coli strain as a model for
highly ATP demanding processes in comparison with an E. coli wildtype
strain. To enable a time-resolved analysis, constant phosphate feeding
strategies were applied to elongate the transition from phosphate
saturated to phosphate limited cell growth. With increasing phosphate
limitation, a reduced cellular efficiency of ATP formation via
respiratory chain activity and the ATP synthase complex was found for
both strains. Process balancing, transcriptome analysis and flux balance
analysis are pointing toward a multi-stage decoupling scenario, which in
essence deteriorates the stoichiometric ratio of ATP formation to proton
translocation, thereby affecting ATP availability from respiration and
carbon usage. Starting off with a potential influence on ATP-synthase
efficiency (stage 1), decoupling is further increased by modified
respiratory activity (stage 2) and by product overflow (stage 3) finally
resulting in a metabolic breakdown entering complete phosphate depletion
(stage 4). The decoupling is initiated by phosphate limitation; further
effects are mainly mediated on metabolic level through ATP availability
and energy charge, additionally affected by ATP demanding product
synthesis. (C) 2014 Elsevier B.V. All rights reserved.
The authors like to thank the company Evonik Industries AG, namely
Thomas Durhuus, Dr. Robert Gerstmeir, Dr. Mechthild Rieping and Dr. Ralf
Kelle as well as the German Funding Agency BMBF for project support
(grant number: 0313917A). Furthermore, the authors like to thank Dr.
Oliver Vielhauer for helpful hints on sample preparation and
metabolomics.
%0 Journal Article
%1 ISI:000345527800011
%A Schuhmacher, Tom
%A Loeffler, Michael
%A Hurler, Thilo
%A Takors, Ralf
%C PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
%D 2014
%I ELSEVIER SCIENCE BV
%J JOURNAL OF BIOTECHNOLOGY
%K myown
%P 96-104
%R 10.1016/j.jbiotec.2014.04.025
%T Phosphate limited fed-batch processes: Impact on carbon usage and energy
metabolism in Escherichia coli
%U https://doi.org/10.1016/j.jbiotec.2014.04.025
%V 190
%X Phosphate starvation is often applied as a tool to limit cell growth in
microbial production processes without hampering carbon and/or nitrogen
supply alternatively. This contribution focuses on the interplay of
process induced phosphate starvation and microbial performance studying
an l-tryptophan overproducing Escherichia coli strain as a model for
highly ATP demanding processes in comparison with an E. coli wildtype
strain. To enable a time-resolved analysis, constant phosphate feeding
strategies were applied to elongate the transition from phosphate
saturated to phosphate limited cell growth. With increasing phosphate
limitation, a reduced cellular efficiency of ATP formation via
respiratory chain activity and the ATP synthase complex was found for
both strains. Process balancing, transcriptome analysis and flux balance
analysis are pointing toward a multi-stage decoupling scenario, which in
essence deteriorates the stoichiometric ratio of ATP formation to proton
translocation, thereby affecting ATP availability from respiration and
carbon usage. Starting off with a potential influence on ATP-synthase
efficiency (stage 1), decoupling is further increased by modified
respiratory activity (stage 2) and by product overflow (stage 3) finally
resulting in a metabolic breakdown entering complete phosphate depletion
(stage 4). The decoupling is initiated by phosphate limitation; further
effects are mainly mediated on metabolic level through ATP availability
and energy charge, additionally affected by ATP demanding product
synthesis. (C) 2014 Elsevier B.V. All rights reserved.
@article{ISI:000345527800011,
abstract = {{Phosphate starvation is often applied as a tool to limit cell growth in
microbial production processes without hampering carbon and/or nitrogen
supply alternatively. This contribution focuses on the interplay of
process induced phosphate starvation and microbial performance studying
an l-tryptophan overproducing Escherichia coli strain as a model for
highly ATP demanding processes in comparison with an E. coli wildtype
strain. To enable a time-resolved analysis, constant phosphate feeding
strategies were applied to elongate the transition from phosphate
saturated to phosphate limited cell growth. With increasing phosphate
limitation, a reduced cellular efficiency of ATP formation via
respiratory chain activity and the ATP synthase complex was found for
both strains. Process balancing, transcriptome analysis and flux balance
analysis are pointing toward a multi-stage decoupling scenario, which in
essence deteriorates the stoichiometric ratio of ATP formation to proton
translocation, thereby affecting ATP availability from respiration and
carbon usage. Starting off with a potential influence on ATP-synthase
efficiency (stage 1), decoupling is further increased by modified
respiratory activity (stage 2) and by product overflow (stage 3) finally
resulting in a metabolic breakdown entering complete phosphate depletion
(stage 4). The decoupling is initiated by phosphate limitation; further
effects are mainly mediated on metabolic level through ATP availability
and energy charge, additionally affected by ATP demanding product
synthesis. (C) 2014 Elsevier B.V. All rights reserved.}},
added-at = {2018-06-08T11:30:24.000+0200},
address = {{PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}},
affiliation = {{Takors, R (Reprint Author), Univ Stuttgart, Inst Biochem Engn, Allmandring 31, D-70569 Stuttgart, Germany.
Schuhmacher, Tom; Loeffler, Michael; Hurler, Thilo; Takors, Ralf, Univ Stuttgart, Inst Biochem Engn, D-70569 Stuttgart, Germany.}},
author = {Schuhmacher, Tom and Loeffler, Michael and Hurler, Thilo and Takors, Ralf},
author-email = {{schuhmacher@ibvt.uni-stuttgart.de
Michael.loeffler@ibvt.uni-stuttgart.de
thilo.hurler@arcor.de
takors@ibvt.uni-stuttgart.de}},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/25a9ed4c7918fd6133c67a35e52697b9e/ralftakors},
da = {{2018-01-26}},
doc-delivery-number = {{AU3QF}},
doi = {{10.1016/j.jbiotec.2014.04.025}},
eissn = {{1873-4863}},
funding-acknowledgement = {{German Funding Agency BMBF {[}0313917A]}},
funding-text = {{The authors like to thank the company Evonik Industries AG, namely
Thomas Durhuus, Dr. Robert Gerstmeir, Dr. Mechthild Rieping and Dr. Ralf
Kelle as well as the German Funding Agency BMBF for project support
(grant number: 0313917A). Furthermore, the authors like to thank Dr.
Oliver Vielhauer for helpful hints on sample preparation and
metabolomics.}},
interhash = {6d83f40fc9790cdb3c1f25bdfcd64b02},
intrahash = {5a9ed4c7918fd6133c67a35e52697b9e},
issn = {{0168-1656}},
journal = {{JOURNAL OF BIOTECHNOLOGY}},
journal-iso = {{J. Biotechnol.}},
keywords = {myown},
keywords-plus = {{PHO REGULON; TRANSCRIPTOME ANALYSIS; STARVATION RESPONSE; ATP SYNTHASE;
P-I; GROWTH; GENE; BACTERIA; OPERON; STOICHIOMETRY}},
language = {{English}},
month = {{NOV 20}},
number-of-cited-references = {{45}},
pages = {{96-104}},
publisher = {{ELSEVIER SCIENCE BV}},
research-areas = {{Biotechnology \& Applied Microbiology}},
times-cited = {{6}},
timestamp = {2018-06-08T09:30:24.000+0200},
title = {{Phosphate limited fed-batch processes: Impact on carbon usage and energy
metabolism in Escherichia coli}},
type = {{Article}},
unique-id = {{ISI:000345527800011}},
url = {https://doi.org/10.1016/j.jbiotec.2014.04.025},
usage-count-last-180-days = {{1}},
usage-count-since-2013 = {{34}},
volume = {{190}},
web-of-science-categories = {{Biotechnology \& Applied Microbiology}},
year = {{2014}}
}