丹参萜类合酶功能及腺毛天然产物研究

	丹参萜类合酶功能及腺毛天然产物研究; Study on the function of terpene synthases and the natural products in the glandular trichomes of Salvia miltiorrhiza Bunge
	罗柳琼
导师	黎胜红刘燕
摘要	Abstract Salvia miltiorrhiza is an important traditional Chinese medicine belonging to the genus Salvia of the family Lamiaceae. This dissertation is composed of four chapters, summarizing the study work on the function of terpene synthases (TPSs) and the natural products in the glandular trichomes of S. miltiorrhiza. Chapter 1 elaborates the functional study of the canditate terpene synthase genes in S. miltiorrhiza. Chapter 2 expounds the de novo production of an inaccessible sesquiterpene (-)-5-epieremophilene by expressing SmSTPS1 in metabolically engineered Escherichia coli. Chapter 3 describes the investigation on the morphology and distribution of glandular trichomes in S. miltiorrhiza as well as the specific natural product in peltate glandular trichomes. Chaptrer 4 reviews the progress of terpene biosynthesis in S. miltiorrhiza. Chapter 1 Functional analysis of terpene synthases in Salvia Miltiorrhiza bunge Sesterterpenoids are a rare subclass of terpenoids, which are derived from geranylfarnesyl diphosphate (GFDP) catalyzed by sesterterpene synthases. Plant sesterterpenoids, which frequently possessed complex and novel structure as well as entensive biological activity, were mainly distributed in Lamiaceae plants and found to be one of the characteristic compounds of the genus Salvia. Since S. miltiorrhiza also belongs to the genus Salvia and its genome and transcriptome sequences are available, we are interested in whether S. Miltiorrhiza harbors sesterterpene synthases. Based on bioinformatic analysis, gene cloning, heterologous expression, and in vivo enzyme assay in combination with GC-MS analysis, the canditate terpene synthase genes from S. Miltiorrhiza were cloned and functionally analyzed. A total of 16 candidate terpene synthase genes were screened from the genome and transcriptome data of S. Miltiorrhiza. The full-length cDNA of 10 candidate genes were obtained by RT-PCR, and cloned into expression vector pCold-TF, namely pCold-TF/SmCPS1, pCold-TF/SmCPS2, pCold-TF/SmCPS3, pCold-TF/SmCPS4, pCold-TF/SmKSL1, pCold-TF/SmSTPS1, pCold-TF/SmSTPS2, pCold-TF/SmTPSa, pCold-TF/SmTPS5 and pCold-TF/SmTPS7. The recombinant proteins of these candidate genes except pCold-TF/SmKSL1 were well soluble by heterologous expression analysis in E. coli. Furthermore, the activities of diterpene synthase were detected in SmCPS1, SmCPS2 and SmCPS4, and sesquiterpene synthase activities were detected in SmSTPS1 and SmSTPS2, which were in accordance with the results in literature. Unfortunately, none of these 9 candidate genes showed sesterterpene synthase activity in our in vivo enzymatic analysis combined with GC-MS analysis, among which any activity was undetectable in SmCPS3, SmTPSa, SmTPS5 and SmTPS7. Further investigation is still in progress. Chapter 2 Heterologous synthesis of (-)-5-epieremophilene by metabolically engineered Escherichia coli (-)-5-Epieremophilene, the epimer of an extremely versatile sesquiterpene (+)-valencene, is an in
	2020-08
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74160
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	罗柳琼. 丹参萜类合酶功能及腺毛天然产物研究, Study on the function of terpene synthases and the natural products in the glandular trichomes of Salvia miltiorrhiza Bunge[D],2020.

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