Integration of induction, system optimization and genetic transformation in Veratrum californicum var. vitro cultures to enhance the production of cyclopamine and veratramine
文献类型： 外文期刊
作者： Zuo, Anqi 1 ; He, Di 1 ; Sun, Chongrui 1 ; Wen, Yashi 1 ; Li, He 1 ; Kou, Chengxi 2 ; Shao, Gaige 3 ; Xue, Zheyong 2 ; Ma, Rui 4 ; Wei, Jia 4 ; Liu, Jingying 1 ; Ma, Pengda 1 ;
作者机构： 1.Northwest A&F Univ, Coll Life Sci, Yangling 712100, Peoples R China
2.Northeast Forestry Univ, Heilongjiang Key Lab Plant Bioact Subst Biosynth &, Harbin 150040, Peoples R China
3.Xian Agr Technol Extens Ctr, Xian, Peoples R China
4.Jilin Acad Agr Sci, Jilin Prov Key Lab Agr Biotechnol, Changchun 130033, Peoples R China
关键词： Veratrum californicum var.; Steroid alkaloid; Efficient production; VnOSCs
期刊名称：PLANT PHYSIOLOGY AND BIOCHEMISTRY （ 影响因子：5.7； 五年影响因子：6.4 ）
ISSN： 0981-9428
年卷期： 2024 年 216 卷
页码：
收录情况： SCI
摘要： Cyclopamine, a compound found in wild Veratrum has shown promising potential as a lead anti-cancer drug by effectively blocking cancer signaling pathways. However, its complex chemical structure poses challenges for artificial synthesis, thus limiting its supply and downstream drug production. This study comprehensively utilizes induction, system optimization, and transgenic technologies to establish an efficient suspension culture system for the high-yield production of cyclopamine and its precursor, veratramine. Experimental results demonstrate that methyl jasmonate (MeJA) effectively promotes the content of veratramine and cyclopamine in Veratrum californicum var. callus tissue, while yeast extract (YE) addition significantly increases cell biomass. The total content of veratramine and cyclopamine reached 0.0638 mg after synergistic treatment of suspension system with these two elicitors. And the content of the two substances was further increased to 0.0827 mg after the optimization by response surface methodology. Subsequently, a genetic transformation system for V. californicum callus was established and a crucial enzyme gene VnOSC1, involved in the steroidal alkaloid biosynthesis pathway, was screened and identified for genetic transformation. Combined suspension culture and synergistic induction system, the total content of the two substances in transgenic suspension system was further increased to 0.1228 mg, representing a 276.69% improvement compared to the initial culture system. This study proposes a complete and effective genetic transformation and cultivation scheme for V. californicum tissue cells, achieving milligram-level production of the anticancer agent cyclopamine and its direct precursor veratramine for the first time. It provides a theoretical basis for the industrial-scale production of these substances.
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