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The <jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> from <jats:italic>Passiflora\u00a0edulis<\/jats:italic> (<jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content>) is <jats:italic>R<\/jats:italic>\u2010selective and is the smallest <jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> known to date. The crystal structures of <jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> and its C\u2010terminal peptide depleted derivative were determined by molecular replacement method using the template structure of a heat stable protein, <jats:styled-content style=\"fixed-case\">SP<\/jats:styled-content>1, from <jats:italic>Populus tremula<\/jats:italic> at 2.8 and 1.8\u00a0\u00c5 resolution, respectively. <jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> belongs to dimeric \u03b1+\u03b2 barrel superfamily consisting of a central \u03b2\u2010barrel in the middle of a dimer. The structure of <jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> complexed with (<jats:italic>R<\/jats:italic>)\u2010mandelonitrile ((<jats:italic>R<\/jats:italic>)\u2010<jats:styled-content style=\"fixed-case\">MAN<\/jats:styled-content>) was also determined. The hydroxyl group of (<jats:italic>R<\/jats:italic>)\u2010<jats:styled-content style=\"fixed-case\">MAN<\/jats:styled-content> forms hydrogen bonds with His8 and Tyr30 in the active site, whereas the nitrile group is oriented toward the carboxyl group of Glu54, unlike other <jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content>s, where it interacts with basic residues typically. The results of mutational analysis indicate that the catalytic dyad of His8\u2010Asn101 is critical for the enzymatic reaction. The length of the hydrogen bond between His\u2010N\u03b41 and Asn101\u2010O\u03b41 is short in the <jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content>\u2010(<jats:italic>R<\/jats:italic>)\u2010<jats:styled-content style=\"fixed-case\">MAN<\/jats:styled-content> complex (~\u00a02.6\u00a0\u00c5), which would increase the basicity of His8 to abstract a proton from the hydroxyl group of (<jats:italic>R<\/jats:italic>)\u2010<jats:styled-content style=\"fixed-case\">MAN<\/jats:styled-content>. The cyanide ion released from the nitrile group abstracts a proton from the protonated His8 to generate a hydrogen cyanide. Thus, the His8 in the active site of <jats:italic>Pe<\/jats:italic><jats:styled-content style=\"fixed-case\">HNL<\/jats:styled-content> acts both as a general acid and a general base in the reaction.<\/jats:p><\/jats:sec><jats:sec><jats:title>Enzymes<\/jats:title><jats:p>\n<jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.chem.qmul.ac.uk\/iubmb\/enzyme\/EC4\/1\/2\/10.html\">EC 4.1.2.10<\/jats:ext-link>\n<\/jats:p><\/jats:sec><jats:sec><jats:title>Database<\/jats:title><jats:p>Structural data are available in <jats:styled-content style=\"fixed-case\">PDB<\/jats:styled-content> database under the accession numbers <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=5XZQ\">5XZQ<\/jats:ext-link>, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=5XZT\">5XZT<\/jats:ext-link>, and <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=5Y02\">5Y02<\/jats:ext-link>.<\/jats:p><\/jats:sec>","DOI":"10.1111\/febs.14339","type":"journal-article","created":{"date-parts":[[2017,11,20]],"date-time":"2017-11-20T15:26:38Z","timestamp":1511191598000},"page":"313-324","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["The crystal structure and catalytic mechanism of hydroxynitrile lyase from passion fruit, <i>Passiflora edulis<\/i>"],"prefix":"10.1111","volume":"285","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4461-8673","authenticated-orcid":false,"given":"Fumihiro","family":"Motojima","sequence":"first","affiliation":[{"name":"Biotechnology Research Center and Department of Biotechnology Toyama 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