蜗牛酶转化人参皂苷Rb_1制备人参稀有皂苷CompoundK的研究

Study on the preparation of rare ginsenoside Compound K by enzymolysis of ginsenoside Rb1 with snailase

目的:确定蜗牛酶转化人参皂苷Rb1制备人参稀有皂苷Compound K(CK)的主要因素,并优选出转化效率高、稳定、适合工业化生产的工艺条件。方法:采用蜗牛酶水解人参皂苷Rb1制备人参稀有皂苷CK,以转化率为指标,通过单因素考察p H值、温度、底物浓度、酶用量、反应时间对转化率的影响,并通过响应面法试验优化制备工艺;采用MS,1H-NMR,13C-NMR鉴定酶解产物。结果:酶解反应的最优条件为温度55℃、p H值5.5醋酸-醋酸钠缓冲液,底物浓度1.0mg/m L,酶与底物质量比1∶1,反应时间36h,在此条件下的转化率为86.91%;经核磁图谱证实产物为人参稀有皂苷CK。结论:蜗牛酶水解人参皂苷Rb1制备人参稀有皂苷CK反应条件温和,工艺简单可靠,适合工业化生产。

Objective： Determination of the main factors which could convert the ginsenoside Rb1 into the rare ginsenoside Compound K by using the snailase, to select the process conditions which were efficient, stable and suitable for the industrial production. Methods： Considering the bioconversion rate as the target, snailase was used for the enzymolysis between the ginsenoside Rb1 and the rare ginsenoside Compund K. Furthermore, the effectiveness of p H value, temperature, concentration, dosage of enzyme and reaction time on the bioconversion rate were studied, and the response surface method was used to optimize the preparation conditions. The hydrolysis product was identified by MS, 1H-NMR, and 13C-NMR. Results： The optimum conditions of enzymatic hydrolysis reaction were achieved at 50℃, p H 5.5 of the acetic acid-sodium acetate（1∶1） buffer solution for 36 h. The substrate concentration was 1.0mg/m L. With the above conditions, the bioconversion rate was 86.91%, NMR spectra confirmed that the product was rare ginsenoside Compund K. Conclusion： It is feasible to prepare rare ginsenoside Compund K by enzymolysis with snailase. The condition is moderate, and the process is simple and reliable, which is suitable for industrialization.