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@ARTICLE{Kappelmann:890651,
author = {Kappelmann, Jannick and Klein, Bianca and Papenfuß,
Mathias and Lange, Julian and Blombach, Bastian and Takors,
Ralf and Wiechert, Wolfgang and Polen, Tino and Noack,
Stephan},
title = {{C}omprehensive {A}nalysis of {C}. glutamicum {A}naplerotic
{D}eletion {M}utants {U}nder {D}efined d-{G}lucose
{C}onditions},
journal = {Frontiers in Bioengineering and Biotechnology},
volume = {8},
issn = {2296-4185},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {FZJ-2021-01099},
pages = {602936},
year = {2021},
abstract = {Wild-type C. glutamicum ATCC 13032 is known to possess two
enzymes with anaplerotic (C4-directed) carboxylation
activity, namely phosphoenolpyruvate carboxylase (PEPCx) and
pyruvate carboxylase (PCx). On the other hand, C3-directed
decarboxylation can be catalyzed by the three enzymes
phosphoenolpyruvate carboxykinase (PEPCk), oxaloacetate
decarboxylase (ODx), and malic enzyme (ME). The resulting
high metabolic flexibility at the anaplerotic node
compromises the unambigous determination of its carbon and
energy flux in C. glutamicum wild type. To circumvent this
problem we performed a comprehensive analysis of selected
single or double deletion mutants in the anaplerosis of
wild-type C. glutamicum under defined D-glucose conditions.
By applying well-controlled lab-scale bioreactor experiments
in combination with untargeted proteomics, quantitative
metabolomics and whole-genome sequencing hitherto unknown,
and sometimes counter-intuitive, genotype-phenotype
relationships in these mutants could be unraveled. In
comparison to the wild type the four mutants C. glutamiucm
Δpyc, C. glutamiucm Δpyc Δodx, C. glutamiucm Δppc Δpyc,
and C. glutamiucm Δpck showed lowered specific growth rates
and D-glucose uptake rates, underlining the importance of
PCx and PEPCk activity for a balanced carbon and energy flux
at the anaplerotic node. Most interestingly, the strain C.
glutamiucm Δppc Δpyc could be evolved to grow on D-glucose
as the only source of carbon and energy, whereas this
combination was previously considered lethal. The prevented
anaplerotic carboxylation activity of PEPCx and PCx was
found in the evolved strain to be compensated by an
up-regulation of the glyoxylate shunt, potentially in
combination with the 2-methylcitrate cycl},
cin = {IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2171 - Biological and environmental resources for
sustainable use (POF4-217)},
pid = {G:(DE-HGF)POF4-2171},
typ = {PUB:(DE-HGF)16},
pubmed = {33553115},
UT = {WOS:000614085700001},
doi = {10.3389/fbioe.2020.602936},
url = {https://juser.fz-juelich.de/record/890651},
}