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Regulatory Mechanisms of Terpene Biosynthesis in Plant Cultured Cells

植物培养细胞萜烯生物合成的调控机制

基本信息

批准号：
03660123
负责人：
NABETA Kensuke
金额：
$ 1.34万
依托单位：
Obihiro University of Agriculture and Veterinary Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

1. A Cyclization of farnesyl pyrophophate (FPP) to bisabolane class sesquiterpenes including alpha-cedrene by cell-free extract from Larix leptolepis callus was investigated. The cell-free extracts specifically converted the 2Z-isomers of FPP to form alpha- cedrene. The cell-free extract was fractionated by centrifugation. The activity to form alpha-cedrene was present in 2000 g supernatant, but absent in 46,200 g supernatant. The activity was solubilized by detergents, but not EDTA. The results suggested that the enzyme is amphipathic or hydrophobic membrane protein. The absolute configuration of alpha-cedrene which is accumulated in Larix leptolepis callus was determined as (+)-alpha- isomer by GC and GC/MS using a chiral capillary column. The occurrence of (+)-alpha-cedrene was confirmed for the first time in nature. These properties of enzymes and a stereochemical course to form (+)-alpha-cedrene were markedly different from those reported for the enzymes from intact plants.2. Lower terpenes produced by liverwort (Heteroscyphus planus) suspension cells were isolated. Structures of (-)-(1S)-7-Methoxy- 1,2-dihydrocadalene, (+)-(1S, 4R)-7-methoxycalamenene (a nobel compound), (+)-(1S,4R)-hydroxycalamenene (a nobel compound), and 7-methoxycadalene were determined by their spectral data and the chemical correlation. Feeding experiments of the specifically deuterated mevalonates indicated several hydride shifts during biosynthesis of cadalene derivatives. A cell-free extract from the suspension cells specifically converted the 6E-isomers of FPP to form sesquiterpenes of cadinane-type.3. The hydride shifts during the formation of bisabolane class compound in Perilla (Akachirimenshiso) callus were confirmed by the feeding experiment using the specifically deuterated mevalonates.

1.研究了日本落叶松愈伤组织无细胞提取物对焦磷酸法呢酯（FPP）环化反应生成α-雪松烯等红没药烷类倍半萜的影响。无细胞提取物特异性地转化FPP的2 Z-异构体以形成α-雪松烯。通过离心分离无细胞提取物。在2000 g上清液中存在形成α-雪松烯的活性，但在46，200 g上清液中不存在。活性被洗涤剂增溶，但不被EDTA增溶。结果表明，该酶为两亲性或疏水性膜蛋白。用GC和GC/MS手性毛细管柱测定了日本落叶松（Larix leptolepis）愈伤组织中α-柏木烯的绝对构型为（+）-α-异构体。（+）-α-雪松烯的存在首次在自然界中得到证实。酶的这些性质和形成（+）-α-雪松烯的立体化学过程与报道的来自完整植物的酶明显不同。对苔类植物悬浮细胞产生的低萜类化合物进行了分离。本文报道了（-）-（1 S）-7-甲氧基-1，2-二氢卡达烯、（+）-（1 S，4 R）-7-甲氧基calamenene（一种诺贝尔化合物）、（+）-（1 S，4 R）-羟基calamenene（一种诺贝尔化合物）和7-甲氧基卡达烯的结构。特定氘代甲羟戊酸盐的饲养实验表明在Cadalene衍生物的生物合成过程中发生了几次氢化物移位。悬浮细胞的无细胞提取物特异性地转化FPP的6 E-异构体形成杜松烷型倍半萜。用特定氘代甲羟戊酸盐进行的饲养实验证实了紫苏愈伤组织中红没药烷类化合物形成过程中的氢化物位移。