中国现代中药2024，Vol.26Issue(2)：316-327,12.DOI:10.13313/j.issn.1673-4890.20231205003
比较蛋白质组学揭示茉莉酸甲酯诱导的雷公藤甲素生物合成
Comparative Proteomics Reveals the Biosynthesis of Methyl Jasmonate-induced Triptolide
摘要
Abstract
Objective:To explore the biosynthetic pathway of triptolide to increase its yield or for heterologous production in synthetic biology.Methods:Suspended cells treated by methyl jasmonate(MeJA)-induced Tripterygium wilfordii were sequenced by label-free comparative proteomics.The functional genes involved in the biosynthesis of triptolide were screened step by step through Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)analysis,combined with gene transcriptional expression analysis and in vivo gene function studies.Results:MeJA-induced samples from different time points were identified to have 376 differentially expressed proteins,including eight cytochrome P450s(CYP450s),three transcription factors and a 2-oxoglutarate dependent dioxygenase(2ODD).Among them,CYP71BE89,CYP82AQ2 and Tw2ODD had similar expression patterns to the known diterpene synthases TwTPS7(v2)and TwTPS27(v2)involved in the cyclization of triptolide precursors,both under MeJA induction and in different plant tissues,suggesting that they may be related to triptolide biosynthesis.Further in vivo RNA-interference(RNAi)functional studies using plant cells verified that interference with the genes CYP71BE89,CYP82AQ2,and Tw2ODD reduced triptolide accumulation by 24.1%,41.4%,and 71.4%,respectively,indicating a direct correlation between the three genes and triptolide biosynthesis.Conclusion:The application of comparative proteomics technology reveals several candidate proteins associated with triptolide biosynthesis.Further characterization of the relationship between the biosynthesis of triptolide and exogenous hormone stimulation may provide an effective method for uncovering the biosynthetic pathways of active ingredients in traditional Chinese medicine.关键词
雷公藤/比较蛋白质组学/雷公藤甲素/细胞色素P450酶/2-氧戊二酸依赖的双加氧酶Key words
Tripterygium wilfordii Hook.f./comparative proteomics/triptolide/cytochrome P450/2-oxoglutarate dependent dioxygenase分类
医药卫生引用本文复制引用
张逸风,赵瑜君,苏平,吴晓毅,夏梦,高伟,黄璐琦..比较蛋白质组学揭示茉莉酸甲酯诱导的雷公藤甲素生物合成[J].中国现代中药,2024,26(2):316-327,12.基金项目
国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-D-202005) （ZYYCXTD-D-202005）
国家自然科学基金项目(82204547) （82204547）
国家重点研发计划项目(2020YFA0908000) （2020YFA0908000）
中国中医科学院科技创新工程项目(CI2023E002) （CI2023E002）
