000023196 001__ 23196 000023196 005__ 20190625110850.0 000023196 0247_ $$2pmid$$apmid:22851196 000023196 0247_ $$2pmc$$apmc:PMC3444685 000023196 0247_ $$2DOI$$a10.1002/cbic.201200239 000023196 0247_ $$2WOS$$aWOS:000308037100014 000023196 0247_ $$2altmetric$$aaltmetric:866944 000023196 037__ $$aPreJuSER-23196 000023196 041__ $$aeng 000023196 082__ $$a540 000023196 084__ $$2WoS$$aBiochemistry & Molecular Biology 000023196 084__ $$2WoS$$aChemistry, Medicinal 000023196 1001_ $$0P:(DE-HGF)0$$aAndexer, J. N.$$b0 000023196 245__ $$aHydroxynitrile lyases with alpha/beta-hydrolase fold: two enzymes with almost identical 3D structures but opposite enantioselectivities and different reaction mechanisms 000023196 260__ $$aWeinheim$$bWiley-VCH$$c2012 000023196 300__ $$a1932 - 1939 000023196 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article 000023196 3367_ $$2DataCite$$aOutput Types/Journal article 000023196 3367_ $$00$$2EndNote$$aJournal Article 000023196 3367_ $$2BibTeX$$aARTICLE 000023196 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000023196 3367_ $$2DRIVER$$aarticle 000023196 440_0 $$08962$$aChemBioChem$$v13$$x1439-4227$$y13 000023196 500__ $$aWe acknowledge financial support from the Austrian Science Foundation (FWF) through projects P17132 (to C.K.) and L148 (to K.G.). X-ray diffraction data were collected at the EMBL/DESY beamline X13 in Hamburg, Germany and we are indebted to the beamline staff for their help. For her help with enzyme purification, we thank Ilona Frindi-Wosch. 000023196 520__ $$aHydroxynitrile 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. 000023196 536__ $$0G:(DE-Juel1)FUEK410$$2G:(DE-HGF)$$aBiotechnologie$$cPBT$$x0 000023196 588__ $$aDataset connected to Web of Science, Pubmed 000023196 65320 $$2Author$$abiocatalysis 000023196 65320 $$2Author$$aC-C lyase 000023196 65320 $$2Author$$aenzyme mechanisms 000023196 65320 $$2Author$$amolecular modeling 000023196 65320 $$2Author$$aX-ray crystallography 000023196 650_7 $$2WoSType$$aJ 000023196 7001_ $$0P:(DE-HGF)0$$aStaunig, N.$$b1 000023196 7001_ $$0P:(DE-HGF)0$$aEggert, T.$$b2 000023196 7001_ $$0P:(DE-HGF)0$$aKratky, C.$$b3 000023196 7001_ $$0P:(DE-Juel1)131522$$aPohl, M.$$b4$$uFZJ 000023196 7001_ $$0P:(DE-HGF)0$$aGruber, K.$$b5 000023196 773__ $$0PERI:(DE-600)2020469-3$$a10.1002/cbic.201200239$$gVol. 13, p. 1932 - 1939$$p1932 - 1939$$q13<1932 - 1939$$tChemBioChem$$v13$$x1439-4227$$y2012 000023196 8567_ $$2Pubmed Central$$uhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3444685 000023196 909CO $$ooai:juser.fz-juelich.de:23196$$pVDB 000023196 915__ $$0StatID:(DE-HGF)0010$$2StatID$$aJCR/ISI refereed 000023196 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR 000023196 915__ $$0StatID:(DE-HGF)0110$$2StatID$$aWoS$$bScience Citation Index 000023196 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded 000023196 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection 000023196 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bThomson Reuters Master Journal List 000023196 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS 000023196 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline 000023196 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database 000023196 915__ $$0StatID:(DE-HGF)1030$$2StatID$$aDBCoverage$$bCurrent Contents - Life Sciences 000023196 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews 000023196 9141_ $$y2012 000023196 9131_ $$0G:(DE-Juel1)FUEK410$$1G:(DE-HGF)POF2-890$$2G:(DE-HGF)POF2-800$$aDE-HGF$$baußerhalb PoF$$kPBT$$lohne FE$$vBiotechnologie$$x0 000023196 9132_ $$0G:(DE-HGF)POF3-581$$1G:(DE-HGF)POF3-580$$2G:(DE-HGF)POF3-500$$aDE-HGF$$bKey Technologies$$lKey Technologies for the Bioeconomy$$vBiotechnology$$x0 000023196 9201_ $$0I:(DE-Juel1)VDB56$$gIBT$$kIBT-2$$lBiotechnologie 2$$x0$$zab 31.10.10 weitergeführt IBG-1 000023196 970__ $$aVDB:(DE-Juel1)140179 000023196 980__ $$aVDB 000023196 980__ $$aConvertedRecord 000023196 980__ $$ajournal 000023196 980__ $$aI:(DE-Juel1)IBG-1-20101118 000023196 980__ $$aUNRESTRICTED 000023196 981__ $$aI:(DE-Juel1)IBG-1-20101118