中草药重要功能成分α-法尼烯检测方法建立  

Establishment of a detection method for α-farnesene

作　　者：郑丹妮 陈思哲 吴芷玥 张一涵 崔瑜 薛闯[1,2] 康巍 ZHENG Danni;CHEN Sizhe;WU Zhiyue;ZHANG Yihan;CUI Yu;XUE Chuang;KANG Wei(School of Bioengineering,Dalian University of Technology,Dalian 116024,China;Ningbo Institute of Dalian University of Technology,Ningbo 315016,China;The Third Hospital of Shijiazhuang,Shijiazhuang 050011,China)

机构地区：[1]大连理工大学生物工程学院,辽宁大连116024 [2]大连理工大学宁波研究院,浙江宁波116038 [3]石家庄第三医院,石家庄050011

出　　处：《吉林中医药》2022年第1期88-91,共4页Jilin Journal of Chinese Medicine

基　　金：国家自然科学基金青年项目(21807043);大连理工大学基本科研业务费项目[DUT20RC(3)069]。

摘　　要：目的α-法尼烯在一系列中草药中发挥着重要作用。本研究将利用生物合成法合成α-法尼烯,并建立一种灵敏、快速检测细胞提取物中α-法尼烯成分的方法。方法通过构建表达α-法尼烯合酶质粒,将其转入大肠杆菌,诱导该酶在细菌内进行表达并利用大肠杆菌MEP途径的代谢产物法尼基焦磷酸(FPP)作为底物,生产α-法尼烯;经正十二烷萃取,以氦气为载气上样,通过气相色谱-质谱联用(GC-MS)方法进行分析。检测9.21 min 出峰物质,对其结构进行质谱分析,确定产物为α-法尼烯。结果细胞提取物中α-法尼烯产物含量为3.63 mg/L。GC-MS可以检测浓度低至50 g/L的α-法尼烯。结论通过GC-MS建立了高效灵敏的α-法尼烯检测方法。Objective To synthesize α-farnesene by biosynthesis and establish a sensitive and rapid method for detecting α-farnesene in cell extracts.Methods The plasmid expressing α-farnesene synthase was constructed and transferred into Escherichia coli.The enzyme was induced to express in the bacteria and farnesene pyrophosphate (FPP),a metabolite of the MEP pathway of Escherichia coli,was used as the substrate to produce α-farnesene.Extracted by n-dodecane,loaded with helium as a carrier gas,and analyzed by gas chromatography-mass spectrometry (GC-MS),the peak substance eluted at 9.21 minute was detected,and its structure was analyzed by mass spectrometry,and the product was identified as α-farnesene.Results The content of α-farnesene product in the cell extract was 3.63 mg/L.GC-MS could detect α-farnesene at a concentration as low as 50 g/L.Conclusion An efficient and sensitive detection method for α-farnesene has been established by GC-MS.

关 键 词：中草药 Α-法尼烯 姜黄 GC-MS α-法尼烯合酶 

分 类 号：R284[医药卫生—中药学]