Purification and structural analysis of volatile sesquiterpenes produced by <i>Escherichia coli</i> carrying unidentified terpene synthase genes from edible plants of the family <i>Araliaceae</i>

<p>A simple method to purify volatile sesquiterpenes from recombinant <i>Escherichia coli</i> was developed using the cells that carried known sesquiterpene synthase (<i>Tps</i>) genes <i>ZzZss</i>2 (<i>ZSS</i>2) and <i>ZoTps</i>1. This method was applied for the purification and structural analyses of volatile sesquiterpenes produced by <i>E. coli</i> cells that carried unidentified <i>Tps</i> genes, which were isolated from the <i>Aralia</i>-genus edible plants belonging to the family <i>Araliaceae</i>. Recombinant cells carrying each Tps gene were cultured in the two-layer medium (<i>n</i>-octane/TB medium), and volatile sesquiterpenes trapped in <i>n</i>-octane were purified through two-phase partition, silica gel column chromatography, and reversed-phase preparative high-performance liquid chromatography, if necessary. Further, their structures were confirmed by nuclear magnetic resonance, [α]<sub>D</sub>, and gas chromatography–mass spectrometry analyses. Herein, the products of <i>E. coli</i> cells that carried two <i>Tps</i> gene (named <i>AcTps</i>1 and <i>AcTps</i>2) in <i>Araria cordata</i> “Udo” and a <i>Tps</i> gene (named <i>AeTps</i>1) in <i>Aralia elata</i> “Taranoki” were studied resulting in identifying functionalities of these cryptic <i>Tps</i> genes.</p> <p>Purification and structural analysis of volatile sesquiterpenes produced by <i>Escherichia coli</i> carrying unidentified terpene synthase genes from edible plants of the family <i>Araliaceae</i>.</p>