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@ARTICLE{Hermann:859976,
author = {Hermann, Johannes and Nowotny, Phillip and Schrader, Tobias
E. and Biggel, Philipp and Hekmat, Dariusch and
Weuster-Botz, Dirk},
title = {{N}eutron and {X}-ray crystal structures of {L}actobacillus
brevis alcohol dehydrogenase reveal new insights into
hydrogen-bonding pathways},
journal = {Acta crystallographica / F Structural biology
communications Section F},
volume = {74},
number = {12},
issn = {2053-230X},
address = {Oxford [u.a.]},
publisher = {Blackwell},
reportid = {FZJ-2019-00782},
pages = {754 - 764},
year = {2018},
abstract = {Lactobacillus brevis alcohol dehydrogenase (LbADH) is a
well studied homotetrameric enzyme which catalyzes the
enantioselective reduction of prochiral ketones to the
corresponding secondary alcohols. LbADH is stable and
enzymatically active at elevated temperatures and accepts a
broad range of substrates, making it a valuable tool in
industrial biocatalysis. Here, the expression, purification
and crystallization of LbADH to generate large, single
crystals with a volume of up to 1 mm3 suitable for neutron
diffraction studies are described. Neutron diffraction data
were collected from an H/D-exchanged LbADH crystal using the
BIODIFF instrument at the Heinz Maier-Leibnitz Zentrum
(MLZ), Garching, Germany to a resolution dmin of 2.15 Å
in 16 days. This allowed the first neutron crystal structure
of LbADH to be determined. The neutron structure revealed
new details of the hydrogen-bonding network originating from
the ion-binding site of LbADH and provided new insights into
the reasons why divalent magnesium (Mg2+) or manganese
(Mn2+) ions are necessary for its activity. X-ray
diffraction data were obtained from the same crystal at the
European Synchrotron Radiation Facility (ESRF), Grenoble,
France to a resolution dmin of 1.48 Å. The
high-resolution X-ray structure suggested partial occupancy
of Mn2+ and Mg2+ at the ion-binding site. This is supported
by the different binding affinity of Mn2+ and Mg2+ to the
tetrameric structure calculated via free-energy
molecular-dynamics simulations.},
cin = {JCNS-FRM-II / Neutronenstreuung ; JCNS-1},
ddc = {530},
cid = {I:(DE-Juel1)JCNS-FRM-II-20110218 /
I:(DE-Juel1)JCNS-1-20110106},
pnm = {6G15 - FRM II / MLZ (POF3-6G15) / 6215 - Soft Matter,
Health and Life Sciences (POF3-621) / 6G4 - Jülich Centre
for Neutron Research (JCNS) (POF3-623)},
pid = {G:(DE-HGF)POF3-6G15 / G:(DE-HGF)POF3-6215 /
G:(DE-HGF)POF3-6G4},
experiment = {EXP:(DE-MLZ)BIODIFF-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30511668},
UT = {WOS:000452221700001},
doi = {10.1107/S2053230X18015273},
url = {https://juser.fz-juelich.de/record/859976},
}