【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。更多还原

【Abstract】 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 submerged 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,5 μm);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),0.42%(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 compounds was carried out.The results showed that DTA negatively correlated with eDTA and PAC,with R2 of 0.857 6 and 0.971 7,respectively,which suggested the role of DTA as an important intermediate in the biosynthesis of triterpenic acids in P.cocos.Conclusion: The sum content of three main terpenoids in submerged cultivation mycelium P.cocos was 5.55 times as that in cultivated material,which strongly suggested the possibility of fermentation in the production of medicinally important triterpenic acids in the future.更多还原