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@ARTICLE{Ramp:909763,
author = {Ramp, Paul and Pfleger, Christopher and Dittrich, Jonas and
Mack, Christina and Gohlke, Holger and Bott, Michael},
title = {{P}hysiological, {B}iochemical, and {S}tructural
{B}ioinformatic {A}nalysis of the {M}ultiple {I}nositol
{D}ehydrogenases from {C}orynebacterium glutamicum},
journal = {Microbiology spectrum},
volume = {10},
number = {5},
issn = {2165-0497},
address = {Birmingham, Ala.},
publisher = {ASM},
reportid = {FZJ-2022-03393},
pages = {e01950-22},
year = {2022},
abstract = {Inositols (cyclohexanehexols) comprise nine isomeric cyclic
sugar alcohols, several of which occur in all domains of
life with various functions. Many bacteria can utilize
inositols as carbon and energy sources via a specific
pathway involving inositol dehydrogenases (IDHs) as the
first step of catabolism. The microbial cell factory
Corynebacterium glutamicum can grow with myo-inositol as a
sole carbon source. Interestingly, this species encodes
seven potential IDHs, raising the question of the reason for
this multiplicity. We therefore investigated the seven IDHs
to determine their function, activity, and selectivity
toward the biologically most important isomers myo-,
scyllo-, and d-chiro-inositol. We created an ΔIDH strain
lacking all seven IDH genes, which could not grow on the
three inositols. scyllo- and d-chiro-inositol were
identified as novel growth substrates of C. glutamicum.
Complementation experiments showed that only four of the
seven IDHs (IolG, OxiB, OxiD, and OxiE) enabled growth of
the ΔIDH strain on two of the three inositols. The kinetics
of the four purified enzymes agreed with the complementation
results. IolG and OxiD are NAD+-dependent IDHs accepting
myo- and d-chiro-inositol but not scyllo-inositol. OxiB is
an NAD+-dependent myo-IDH with a weak activity also for
scyllo-inositol but not for d-chiro-inositol. OxiE on the
other hand is an NAD+-dependent scyllo-IDH showing also good
activity for myo-inositol and a very weak activity for
d-chiro-inositol. Structural models, molecular docking
experiments, and sequence alignments enabled the
identification of the substrate binding sites of the active
IDHs and of residues allowing predictions on the substrate
specificity. IMPORTANCE myo-, scyllo-, and d-chiro-inositol
are C6 cyclic sugar alcohols with various biological
functions, which also serve as carbon sources for microbes.
Inositol catabolism starts with an oxidation to
keto-inositols catalyzed by inositol dehydrogenases (IDHs).
The soil bacterium C. glutamicum encodes seven potential
IDHs. Using a combination of microbiological, biochemical,
and modeling approaches, we analyzed the function of these
enzymes and identified four IDHs involved in the catabolism
of inositols. They possess distinct substrate preferences
for the three isomers, and modeling and sequence alignments
allowed the identification of residues important for
substrate specificity. Our results expand the knowledge of
bacterial inositol metabolism and provide an important basis
for the rational development of producer strains for these
valuable inositols, which show pharmacological activities
against, e.g., Alzheimer's disease, polycystic ovarian
syndrome, or type II diabetes. Keywords: Corynebacterium
glutamicum; d-chiro-inositol; enzyme kinetics; inositol
dehydrogenase; inositol metabolism; inositols; molecular
docking; myo-inositol; scyllo-inositol; structural models.},
cin = {IBG-4 / JSC / NIC / IBI-7 / IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IBG-4-20200403 / I:(DE-Juel1)JSC-20090406 /
I:(DE-Juel1)NIC-20090406 / I:(DE-Juel1)IBI-7-20200312 /
I:(DE-Juel1)IBG-1-20101118},
pnm = {5111 - Domain-Specific Simulation $\&$ Data Life Cycle Labs
(SDLs) and Research Groups (POF4-511) / 5241 - Molecular
Information Processing in Cellular Systems (POF4-524) / 2171
- Biological and environmental resources for sustainable use
(POF4-217) / Forschergruppe Gohlke $(hkf7_20200501)$ / BioSC
- Bioeconomy Science Center (BioSC) /
$BioökonomieREVIER_INNO:$ Entwicklung der Modellregion
BioökonomieREVIER Rheinland, TP A (031B0918A)},
pid = {G:(DE-HGF)POF4-5111 / G:(DE-HGF)POF4-5241 /
G:(DE-HGF)POF4-2171 / $G:(DE-Juel1)hkf7_20200501$ /
G:(DE-Juel1)BioSC / G:(BMBF)031B0918A},
typ = {PUB:(DE-HGF)16},
pubmed = {36094194},
UT = {WOS:000854191500002},
doi = {10.1128/spectrum.01950-22},
url = {https://juser.fz-juelich.de/record/909763},
}