人参皂苷Rh_2葡萄糖基转移酶活性测定体系的建立

Establishment of Measurement System of Activity of Ginsenoside Rh_2 Glycosyltransferase

[目的]建立人参细胞中尿苷二磷酸葡萄糖(UDPG):人参皂苷Rh2葡萄糖基转移酶(UGRh2GT)活性测定体系。[方法]以人参皂苷Rh2为底物,以UDPG为葡萄糖供体,选取合适的温度和pH值,用提取的人参细胞酶液催化Rh2生成Rg3,采用高效液相色谱法(HPLC)测定Rg3的生成量,色谱柱为C18柱(150mm×4.6mm),流动相为乙腈和水,梯度洗脱。[结果]人参细胞中UGRh2GT活性测定体系的最优pH值是9.3,最优温度是34.1℃。[结论]建立的酶活测定体系能够较好较方便地测定UGRh2GT的比活。

[Objective] The aim was to establish activity assay system of uridine 5′-diphosphoglucose （UDPG）： ginsenoside Rh2 Glycosyltransferase （UGRh2GT） of ginseng cells. [Method] With UDPG as the glucose donor, Ginsenoside Rh2 has been catalyzed to Rg3 by enzymes which have been extracted from ginseng suspension cells at appropriate temperature and pH. The amount of generated ginsenoside Rg3 has been determined by high performance liquid chromatography （HPLC）.The analytical column was C18 （150 × 4.6 mmol/L）.Mobile phases were acetonitrile and water with gradient elution. [Result] In the activity assay system of UGRh2GT of ginseng cells, the optimum pH value is 9.3, the optimal temperature is 34.1 ℃. [Conclusion] The method of activity assay can determine the activity of UGRh2GT more easily.