Terreusinones类生物碱的生物合成研究

Terreusinones是从真菌Penicillium dangeardii等中分离得到的一类具有蛋白酪氨酸磷酸酶抑制活性的生物碱类化合物，其独特的双吡咯并苯醌的三环骨架预示着独特的生物合成机制。申请人前期经生物信息学分析、化学喂养、基因敲除实验等确定了terreusinone的生物合成基因簇，为本项研究奠定基础。拟利用构巢曲霉异源表达、基因敲除、化学回补、蛋白表达及体内外生化实验从基因-蛋白水平揭示terreusinone的生物合成途径，重点研究双吡咯并苯醌的形成机制及特殊氧化酶的催化机制，并利用合成生物学的策略获取terreusinone的类似物，实现其结构优化。本项研究有望发现催化双吡咯并苯醌形成的特殊酶和新的催化机制，为terreusinones类天然产物的合成与结构优化开辟新的途径，并有望发现潜在的新药先导化合物。

Terreusinones, a kind of novel dipyrrolobenzoquinone alkaloids, has been isolated as protein tyrosine phosphatases inhibitors from endophytic fungus Penicillium dangeardii Pitt. Due to their novel structural framework and significant bioactivities, they have inspired numerous chemical and biologic studies. However, the genetic and biochemical basis of terreusinone biosynthesis has remained unexplored. Herein, we try to identify and characterize of the terreusinone (ter) biosynthetic gene cluster from Penicillium dangeardii. To verify the ter gene cluster and elucidate the biosynthetic pathway of terreusinone, heterologous expression experiments in Aspergillus nidulans, gene knockout, chemical complement, protein expression and biochemical reaction were conducted. The study on the biosynthesis of terreusinone is expected to discover some remarkable enzymes with special functions and bioactive analogs.