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@ARTICLE{Marienhagen:950,
author = {Marienhagen, J. and Eggeling, L.},
title = {{M}etabolic {F}unction of {C}orynebacterium glutamicum
{A}minotransferases {A}la{T} and {A}vt{A} and {I}mpact on
{L}-{V}aline {P}roduction},
journal = {Applied and environmental microbiology},
volume = {74},
issn = {0099-2240},
address = {Washington, DC [u.a.]},
publisher = {Soc.},
reportid = {PreJuSER-950},
pages = {7457 - 7462},
year = {2008},
note = {We thank H. Sahm and M. Bott for continuous support, K.
Krumbach for strain construction, and T. Bartek and C.
Rudolf (IBT 2, Forschungszentrum Julich, Germany) for
assistance with the SIXFORS fermentations.This work was
supported by Deutsche Bundestiftung Umwelt Projekt 13158.},
abstract = {Aminotransferases (ATs) interacting with L-alanine are the
least studied bacterial ATs. Whereas AlaT converts pyruvate
to L-alanine in a glutamate-dependent reaction, AvtA is able
to convert pyruvate to L-alanine in an L-valine-dependent
manner. We show here that the wild type of Corynebacterium
glutamicum with a deletion of either of the corresponding
genes does not exhibit an explicit growth deficiency.
However, a double mutant was auxotrophic for L-alanine,
showing that both ATs can provide L-alanine and that they
are the only ATs involved. Kinetic studies with isolated
enzymes demonstrate that the catalytic efficiency,
k(cat)/K(m), of AlaT is higher than 1 order of magnitude in
the direction of L-alanine formation (3.5 x 10(4) M(-1)
s(-1)), but no preference was apparent for AvtA, suggesting
that AlaT is the principal L-alanine-supplying enzyme. This
is in line with the cytosolic L-alanine concentration, which
is reduced in the exponential growth phase from 95 mM to 18
mM by a deletion of alaT, whereas avtA deletion decreases
the L-alanine concentration only to 76 mM. The combined data
show that the presence of both ATs has subtle but obvious
consequences on balancing intracellular amino acid pools in
the wild type. The consequences are more obvious in an
L-valine production strain where a high intracellular
drain-off of the L-alanine precursor pyruvate prevails. We
therefore used deletion of alaT to successfully reduce the
contaminating L-alanine in extracellular accumulated
L-valine by $80\%.$},
keywords = {Alanine: metabolism / Corynebacterium glutamicum:
enzymology / Corynebacterium glutamicum: genetics /
Corynebacterium glutamicum: growth $\&$ development /
Cytosol: chemistry / Gene Deletion / Kinetics / Pyruvic
Acid: metabolism / Substrate Specificity / Transaminases:
genetics / Transaminases: isolation $\&$ purification /
Transaminases: metabolism / Valine: biosynthesis / Pyruvic
Acid (NLM Chemicals) / Alanine (NLM Chemicals) / Valine (NLM
Chemicals) / Transaminases (NLM Chemicals) / J (WoSType)},
cin = {IBT-1},
ddc = {570},
cid = {I:(DE-Juel1)VDB55},
pnm = {Biotechnologie},
pid = {G:(DE-Juel1)FUEK410},
shelfmark = {Biotechnology $\&$ Applied Microbiology / Microbiology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:18931286},
pmc = {pmc:PMC2607172},
UT = {WOS:000261513700001},
doi = {10.1128/AEM.01025-08},
url = {https://juser.fz-juelich.de/record/950},
}