缬草-4,7（11）-二烯在大肠杆菌中的生物合成

doi:10.13560/j.cnki.biotech.bull.1985.2017.01.014

摘要/Abstract

摘要： 缬草-4，7（11）-二烯是一种双环倍半萜烯类化合物，具有镇静、减缓压力、抗焦虑的作用，可用来治疗注意力缺陷多动症（ADHD）等精神疾病。旨在实现缬草-4，7（11）-二烯在大肠杆菌中的从头合成，通过在大肠杆菌BL21（DE3）过表达酵母源，甲羟戊酸途径中催化乙酰辅酶A合成焦磷酸法尼基（FPP）的8个相关基因及来源于缬草属植物（Valeriana officinalis）的缬草-4，7（11）-二烯合酶（VoTPS）基因;并采用水相和有机相双相发酵的方法，通过GC-MS在有机相中检测到了缬草-4，7（11）-二烯，实现了大肠杆菌中从头合成缬草-4，7（11）-二烯。并对IPTG浓度、蛋白诱导温度和选择不同的有机相等方面进行初步优化后，确定发酵条件为：蛋白诱导温度20℃、IPTG的浓度为0.1 mmol/L，有机相为正十二烷。

关键词: 大肠杆菌, 缬草-4, 7（11）-二烯, 生物合成, 缬草-4, 7（11）-二烯合酶（VoTPS）

Abstract: Valerena-4,7（11）-diene is a bicyclic sesquiterpene and shows particularly profound sedative,stress reducing and anxiolytic effects,and it could be used to treat attention deficit hyperactivity disorder（ADHD）and other mental illness. To achieve the de novo biosynthesis of valerena-4,7（11）-diene in Escherichia coli,eight genes of Saccharomyces cerevisiae involved in converting acetyl-CoA into farnesyl diphosphate（FPP）from mevalonate pathway and the valerena-4,7（11）-diene synthase（VoTPS）gene from Valeriana officinalis were co-overexpressed in BL21（DE3）. Adopting the bipolar fermentation method of water phase and organic phase,the valerian-4,7（11）-diene was detected in organic phase by GC-MS,indicating that the synthesis of valerian-4,7（11）-diene was achieved initially in theE. coli. After primarily optimizing the IPTG concentration,the temperature of protein induction and choice of varied organic phases,the fermentation conditions were determined as follows：protein induced temperature was 20℃,the concentration of IPTG was 0.1 mmol/L,and organic phase was dodecane.

Key words: Escherichia coli, valerian-4,7（11）-diene, biosynthesis, valerian-4,7（11）-diene synthase（VoTPS）

刘畅，殷华，庄以彬，刘涛. 缬草-4,7（11）-二烯在大肠杆菌中的生物合成[J]. 生物技术通报, 2017, 33(1): 129-134.

LIU Chang, YIN Hua, ZHUANG Yi-bin, LIU Tao. Biosynthesis of Valerena-4,7（11）-diene in an Engineered Escherichia coli Strain[J]. Biotechnology Bulletin, 2017, 33(1): 129-134.

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