University of British Columbia, Kelowna, BC, Canada.
Planta. 2013 Nov;238(5):983-9. doi: 10.1007/s00425-013-1937-6. Epub 2013 Aug 6.
The essential oil (EO) of Lavandula is dominated by monoterpenes, but can also contain small amounts of sesquiterpenes, depending on species and environmental conditions. For example, the sesquiterpene 9-epi-caryophyllene can make up to 8 % of the EO in a few species, including those commercially propagated for EO production. Here, we report the cloning and functional characterization of 9-epi-caryophyllene synthase (LiCPS) from the glandular trichomes of Lavandula x intermedia, cv. Grosso. The 1,617 bp open reading frame of LiCPS, which did not encode a transit peptide, was expressed in Escherichia coli and the recombinant protein purified by Ni-NTA agarose affinity chromatography. The ca. 60 kDa recombinant protein specifically converted farnesyl diphosphate to 9-epi-caryophyllene. LiCPS also produced a few monoterpenes when assayed with the monoterpene precursor geranyl diphosphate (GPP), but--unlike most monoterpene synthases--was not able to derive detectable amounts of any products from the cis isomer of GPP, neryl diphosphate. The LiCPS transcripts accumulated in developing L. x intermedia flowers and were highly enriched in glandular trichomes, but were not detected in leaves suggesting that the transcriptional expression of this gene is spatially and developmentally regulated.
薰衣草的精油(EO)主要由单萜组成,但也可以包含少量的倍半萜,具体取决于物种和环境条件。例如,倍半萜 9-表石竹烯在包括那些为精油生产而商业繁殖的几个物种中可占精油的 8%。在这里,我们报告了从 Lavandula x intermedia,cv. Grosso 的腺毛中克隆和功能表征 9-表石竹烯合酶(LiCPS)。LiCPS 的 1617bp 开放阅读框未编码转运肽,在大肠杆菌中表达并通过 Ni-NTA 琼脂糖亲和层析纯化重组蛋白。该约 60 kDa 的重组蛋白特异性地将法呢基二磷酸转化为 9-表石竹烯。当用单萜前体香叶基二磷酸(GPP)进行测定时,LiCPS 还产生了一些单萜,但与大多数单萜合酶不同,它不能从 GPP 的顺式异构体,橙花基二磷酸中获得可检测量的任何产物。LiCPS 的转录物在发育中的 L. x intermedia 花朵中积累,并在腺毛中高度富集,但在叶片中未检测到,这表明该基因的转录表达受到空间和发育的调节。