TY  - JOUR
AU  - Hermann, Johannes
AU  - Nowotny, Phillip
AU  - Schrader, Tobias E.
AU  - Biggel, Philipp
AU  - Hekmat, Dariusch
AU  - Weuster-Botz, Dirk
TI  - Neutron and X-ray crystal structures of Lactobacillus brevis alcohol dehydrogenase reveal new insights into hydrogen-bonding pathways
JO  - Acta crystallographica / F Structural biology communications Section F
VL  - 74
IS  - 12
SN  - 2053-230X
CY  - Oxford [u.a.]
PB  - Blackwell
M1  - FZJ-2019-00782
SP  - 754 - 764
PY  - 2018
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:30511668
UR  - <Go to ISI:>//WOS:000452221700001
DO  - DOI:10.1107/S2053230X18015273
UR  - https://juser.fz-juelich.de/record/859976
ER  -