茯苓发酵菌丝体中3种主要三萜酸类成分积累动态研究  

Dynamic accumulation of three main triterpenic acids in submerged cultivation mycelium of Poria cocos

作　　者：张杨[1] 胡高升[1] 韩志福[1] 萧伟[2] 王振东[2] 毕宇安[2] 贾景明[1] 

机构地区：[1]沈阳药科大学中药学院,辽宁沈阳110016 [2]江苏康缘药业股份有限公司药物研究所,江苏连云港222001

出　　处：《中国中药杂志》2013年第9期1355-1359,共5页China Journal of Chinese Materia Medica

摘　　要：目的:考察茯苓发酵菌丝体中3种主要三萜酸类成分动态积累变化。方法:建立茯苓菌的液体培养方法,采用RP-HPLC测定茯苓发酵菌丝体中去氢土莫酸(DTA),3-表去氢土莫酸(eDTA)和猪苓酸C(PAC)3种主要三萜酸类成分的含量,色谱条件如下:PLATISIL ODS色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.5%磷酸水(80∶20);流速1.0 mL.min-1;检测波长242 nm。结果:在培养后的第8天,生物量达到最大值,但是3种主要三萜酸类成分(DTA,eDTA,PAC)的含量在培养周期内呈持续上升趋势,第17天测得3种成分质量分数分别为1.2%(DTA),0.4%(eDTA),1.0%(PAC),均显著高于栽培茯苓中对应成分[0.2%(DTA),0.12%(eDTA),0.16%(PAC)]。另外,3种成分含量比例的线性回归分析结果表明,DTA与eDTA和PAC的含量比例呈显著负相关,相关系数(R2)分别为0.858 7,0.971 7。该结果表明DTA为茯苓中三萜酸类成分生合成途径中的重要中间体。结论:发酵培养17 d的茯苓菌丝体中DTA,eDTA,PAC含量之和为栽培茯苓中含量的5.55倍,说明在本发酵培养条件下,发酵培养生产茯苓中三萜酸类有效成分技术可行。Objective： To study the dymamic accumulation of triterpenic acids production in submerged cultivation mycelium of Poria cocos. Method： Liquid culture method of P. cocos was established and RP-HPLC was applied to determine the contents of three main triterpenic acids dehydrotumulosic acid （DTA）, 3-epi-dehydrotumulosic acid （eDTA） and polyporenic acid C （PAC） in sub- merged cultivation mycelium P. cocos at different culture stages and the contents were compared with cultivated P. cocos. The HPLC method is as follows, column： Plastisil ODS （4.6 mm ×250 mm,5μm） ; mobile phase： ACN/0. 5% phosphate （80： 20） ; flow rate： 1.0 mL · min^-1 ; detective wavelength： 242 nm. Result： The maximum biomass occurred at the 8th d after inoluctation, however, the contents and yield of three compounds increased till the 17th day. The contents of three compounds were 1.2% （ DTA）, O. 42% （eDTA） and 1.0% （PAC） at the 17th day after inoculation, which were significantly higher than that in cultivated material [0.2% （ DTA）, 0.12（eDTA） and 0.16% （PAC） ]. Furthermore, a correlation analysis between the content ratios of three independent com- pounds was carried out. The results showed that DTA negatively correlated with eDTA and PAC, with R2 of 0. 857 6 and 0. 971 7, re- spectively, which suggested the role of DTA as an important intermediate in the biosynthesis of triterpenic acids in P. cocos. Conclu- sion ： The sum content of three main terpenoids in submerged cultivation mycelium P. cocos was 5.55 times as that in cultivated materi- al, which strongly suggested the possibility of fermentation in the production of medicinally important triterpenic acids in the future.

关 键 词：茯苓 发酵 去氢土莫酸 3-表-去氢土莫酸 猪苓酸C RP-HPLC 含量测定 

分 类 号：R284[医药卫生—中药学]