玄参中β-葡萄糖苷酶活性测定条件及干燥过程中酶活性变化的研究  

Investigate optimum conditions and determinate changes of β-glucosidase activity in Scrophularia root under different drying conditions

作　　者：喻欢欢[1] 吉光见稚代 胥秀英[1] 郑一敏[1] 李隆云[2] 

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]重庆市中药研究院,重庆400065

出　　处：《中国中药杂志》2017年第2期274-279,共6页China Journal of Chinese Materia Medica

基　　金：国家科技支撑计划项目(2011BAI13B02-2);重庆市科技研发基地项目(cstc2014ptyjd10001);科技富民强县专项行动计划项目(国科发农[2014]160号);重庆市基本科研业务项目(2015 cstc-jbky-01903)

摘　　要：采用p NPG法测定玄参中β-葡萄糖苷酶的活性,通过单因素及均匀设计实验对提取液的种类、反应体系、反应时间、温度、以及底物浓度等提取条件与反应条件进行了筛选,并在此基础上测定了新鲜玄参在40~100℃干燥温度中酶活性的变化。结果显示,柠檬酸磷酸缓冲液提取效果较好,50℃环境下反应30 min时酶促反应产生的吸光度最大,均匀设计实验确定最佳提取液pH 7.0、酶反应体系pH 6.0、底物浓度20 mmol·L^(-1)。酶活性变化受干燥温度和失水率影响,60~100℃干燥过程中温度升高,随着失水率加快酶活性迅速下降,而40,50℃干燥过程中一定的酶活性始终存在。该研究为环烯醚萜苷类水解机制和最佳干燥工艺的研究奠定了理论基础。To explore the optimum conditions of β-glucosidase activity in Scrophularia root by using p NPG method. The extraction conditions and reaction conditions（ such as extraction liquid type,reaction system,reaction time,temperature,and substrate concentration） were screened by using monofactorial experiment and homogeneous design. Then the changes of β-glucosidase activity in Scrophularia root were detected at the drying temperature of 40-100 ℃. The results showed that citric acid phosphate buffer had better extraction effect,and the maximum absorbance produced by enzymatic reaction was present at 50 ℃ environment after reaction for 30 min. Homogeneous design experiment determined that the optimal conditions were as follows： optimal extraction liquid pH 7. 0; enzymatic reaction system pH 6. 0; substrate concentration 20 mmol· L-1. The change of enzyme activity was affected by drying temperature and water loss rate. In the drying temperature of 60-100 ℃,the enzyme activity was reduced rapidly with the increase in water loss rate,while the activity was seen even with 0 % of water at 40 and 50 ℃. This study has laid the theoretical foundation for research of hydrolysis mechanism of iridoid glycosides and optimum drying process.

关 键 词：玄参 Β-葡萄糖苷酶 提取条件 活性测定条件 

分 类 号：R282.4[医药卫生—中药学]