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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 investigate the dynamic accumulation of three main triterpenoid acids in the mycelium of Poria cocos fermentation. Methods: The liquid culture method of Poria cocos by bacteria was established. The contents of DTA, 3-epi-eDTA and C (PAC) 3 in mycelium of Poria cocos The main triterpene acids were as follows: PLATISIL ODS column (4.6 mm × 250 mm, 5 μm); mobile phase acetonitrile-0.5% phosphoric acid water (80:20); flow rate 1.0 mL · min- Detection wavelength of 242 nm. Results: The biomass reached the maximum on the 8th day after culture, but the content of three main triterpenoid acids (DTA, eDTA, PAC) kept increasing during the cultivation period. On the 17th day, three kinds of biomass The contents of DTA, 0.4% (eDTA) and 1.0% (PAC) were significantly higher than those of the corresponding components [0.2% (DTA), 0.12% (eDTA), 0.16% ]. In addition, the results of linear regression analysis showed that the contents of DTA, eTA and PAC were negatively correlated with the correlation coefficients (R2) of 0.858 7 and 0.971 7, respectively. This result shows that DTA is an important intermediate in the tilanolic acid component synthesis pathway of Poria. Conclusion: The content of DTA, eDTA and PAC in the Poria cocos culture mycelium cultured for 17 days is 5.55 times of the content of Poria cocos culture, which indicates that the technology of fermenting and producing triterpene acid active ingredients in Poria cocos under the condition of fermentation is feasible .