本氏烟草中异源合成抗癌药物葫芦素前体

葫芦素具有显著的抗癌活性，解析其生物合成途径对异源生产葫芦素具有重要意义。葫芦素首先由葫芦二烯醇（Cuol）在氧化鲨烯环化酶（OSCs）家族的葫芦二烯醇合酶（CBS）催化下生成2,3-氧化鲨烯，后者在细胞色素P450(CYP)等酶催化下氧化修饰产生葫芦素系列化合物。本研究在建立高效的根癌农杆菌（Agrobacterium tumefaciens）介导的瞬时表达体系的基础上，将雪胆（Hemsleya chinensis Cogn.）CBS基因（HcOSC6）与截短的燕麦（Avena strigosa）3-羟基-3-甲基-戊二酰辅酶A还原酶基因（tHMGR）共表达，通过不断优化烟草共表达体系，将本氏烟草（Nicotiana benthamiana）叶片中的Cuol产量从2.832 mg/g（干重）提高到9.48 mg/g（干重）。同时，将雪胆的HcCYP87D20共渗入烟草叶片，获得葫芦素生物合成的重要中间体11-羰基-20β-羟基-葫芦二烯醇。本研究结果将为葫芦素的高效异源合成提供理论依据。

Heterologous Synthesis the Precursors of Anticancer Drug Cucurbitacins in Nicotiana benthamiana

The elucidation of cucurbitacin biosynthesis pathway is essential for heterologous production due to its significant anticancer activity. The biosynthesis of cucurbitacin starts from cucurbitadienol (Cuol) catalyzed by cucurbitadienol synthase (CBS) of the oxysqualene cyclase (OSCs) family to form 2,3-oxysqualene which subsequently catalyzed by cytochrome P450 (CYP) and other enzymes for oxidative modification to produce various cucurbitacins. In this study, an efficient transient expression system mediated by Agrobacterium tumefaciens was established. The CBS gene (HcOSC6) from Hemsleya chinensis and truncated 3-hydroxy-3-methyl-glutaryl coenzyme A reductase gene (tHMGR) from Avena strigose were continuously optimized in the tobacco co-expression system to increase Cuol yield from 2.832 mg/g (DW) to 9.48 mg/g (DW) in Nicotiana benthamiana leaves. Meanwhile, HcCYP87D20 of H. chinensis was also co-infiltrated into tobacco leaves to obtain 11-carbonyl-20β-hydroxy-cucurbitol. This study would provide a theoretical basis for efficient heterologous production of cucurbitacin.