洋虫内产β-葡萄糖苷酶内生菌发酵液转化人参皂苷产物分析及其抗肿瘤活性研究

为提高人参皂苷生物利用度，明确人参皂苷药用洋虫体内生物转化规律及其代谢产物抗肿瘤活性，通过MRS（含七叶苷和柠檬酸铁）培养基筛选洋虫内生菌中高产β-葡萄糖苷酶的菌株，制备复合菌发酵剂液体转化人参皂苷提取物，利用高效液相色谱-串联质谱法分析发酵产物中人参皂苷的化学组成及其动态变化规律，采用四甲基噻唑蓝（methylthiazoletrazolium，MTT）染色法阐明发酵产物低（0.5 mg·L^-1）、中（2 mg·L^-1）、高（5 mg·L^-1）剂量对A549细胞抗肿瘤活性的影响。人参提取物中共检测出Re、Rg1、Rf、Rb1、Rc、Rb2、Rd 7种皂苷，代谢产物中有去糖基化人参皂苷Rh1、Rg3和compoud K产生，发酵产物MTT试验显示高剂量组（5 mg·L^-1）反应72 h抑制肿瘤细胞率可达85%。由此表明，利用洋虫内生菌复合菌液转化人参皂苷提取物可提高中药人参和洋虫的药用价值，为中草药的发酵改性提供了新思路。

Fermentation Products Analysis of Producing β-glucosidase Endophytic Bacteria in Martianus dermestoides Transformated to Ginsenoside and Its Anti-tumor Activity

In order to improve the bioavailability of ginsenosides， and clarify the transformated products of ginsenosides in Martianus dermestoides and the anti-tumor activity of their metabolites， the functional strains of high-yield β-glucosidase in the endophytes of Martianus dermestoides were screened by MRS （esculin， ferric citrate） medium to prepare compound fermentation agent liquid fermentation ginsenoside extract. The chemical composition and dynamic changes of ginsenosides in fermentation products were analyzed by HPLC-MS/MS. The anti-tumor activities of the fermentation product in low （0.5 mg·L^-1）， medium （2 mg·L^-1）， and high （5 mg·L^-1） dose groups on A549 cells were clarified using methylthiazoletrazolium （MTT） blue staining method. A total of 7 saponins （Re， Rg1， Rf， Rb1， Rc， Rb2， Rd） were detected in ginseng extract. Among the metabolites， deglycosylated ginsenosides Rh1， Rg3 and compoud K were formed. The MTT experiment showed that the inhibition rate of tumor cells was 85% after treated 72 h in the high does group with the does 5 mg·L^-1. These results suggested that a variety of high-activity stimulating ginsenosides were transformed by endophytic bacteria of Martianus dermestoides， which improved the medicinal value of Chinese herbal medicine ginseng and onion， and provided a new idea for fermentation and modification of Chinese herbal medicine.