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@ARTICLE{Andexer:23196,
author = {Andexer, J. N. and Staunig, N. and Eggert, T. and Kratky,
C. and Pohl, M. and Gruber, K.},
title = {{H}ydroxynitrile lyases with alpha/beta-hydrolase fold: two
enzymes with almost identical 3{D} structures but opposite
enantioselectivities and different reaction mechanisms},
journal = {ChemBioChem},
volume = {13},
issn = {1439-4227},
address = {Weinheim},
publisher = {Wiley-VCH},
reportid = {PreJuSER-23196},
pages = {1932 - 1939},
year = {2012},
note = {We acknowledge financial support from the Austrian Science
Foundation (FWF) through projects P17132 (to C.K.) and L148
(to K.G.). X-ray diffraction data were collected at the
EMBL/DESY beamline X13 in Hamburg, Germany and we are
indebted to the beamline staff for their help. For her help
with enzyme purification, we thank Ilona Frindi-Wosch.},
abstract = {Hydroxynitrile lyases (HNLs) catalyze the cleavage of
cyanohydrins to yield hydrocyanic acid (HCN) and the
respective carbonyl compound and are key enzymes in the
process of cyanogenesis in plants. In organic syntheses,
HNLs are used as biocatalysts for the formation of
enantiopure cyanohydrins. We determined the structure of the
recently identified, R-selective HNL from Arabidopsis
thaliana (AtHNL) at a crystallographic resolution of 2.5 Å.
The structure exhibits an α/β-hydrolase fold, very similar
to the homologous, but S-selective, HNL from Hevea
brasiliensis (HbHNL). The similarities also extend to the
active sites of these enzymes, with a Ser-His-Asp catalytic
triad present in all three cases. In order to elucidate the
mode of substrate binding and to understand the unexpected
opposite enantioselectivity of AtHNL, complexes of the
enzyme with both (R)- and (S)-mandelonitrile were modeled
using molecular docking simulations. Compared to the complex
of HbHNL with (S)-mandelonitrile, the calculations produced
an approximate mirror image binding mode of the substrate
with the phenyl rings located at very similar positions, but
with the cyano groups pointing in opposite directions. A
catalytic mechanism for AtHNL is proposed, in which His236
from the catalytic triad acts as a general base and the
emerging negative charge on the cyano group is stabilized by
main-chain amide groups and an α-helix dipole very similar
to α/β-hydrolases. This mechanistic proposal is
additionally supported by mutagenesis studies.},
keywords = {J (WoSType)},
cin = {IBT-2},
ddc = {540},
cid = {I:(DE-Juel1)VDB56},
pnm = {Biotechnologie},
pid = {G:(DE-Juel1)FUEK410},
shelfmark = {Biochemistry $\&$ Molecular Biology / Chemistry, Medicinal},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:22851196},
pmc = {pmc:PMC3444685},
UT = {WOS:000308037100014},
doi = {10.1002/cbic.201200239},
url = {https://juser.fz-juelich.de/record/23196},
}