黄芩茎叶葡萄糖醛酸水解酶提取工艺、酶学性质、实际应用研究  

Extraction process,enzymatic properties and practical application of glucuronic hydrolase in Scutellaria baicalensis stems and leaves

作　　者：程雨婕 陈旭 刘云华[2] 黄志芳[2] 陈燕[2] 刘玉红[2] 易进海 CHENG Yu-jie;CHEN Xu;LIU Yun-hua;HUANG Zhi-fang;CHEN Yan;LIU Yu-hong;YI Jin-hai(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611130,China;Sichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia Medica,Sichuan Provincial Academy of Chinese Medicine Sciences,Chengdu 610041,China)

机构地区：[1]成都中医药大学药学院,四川成都611130 [2]四川省中医药科学院,中药材品质及创新中药研究四川省重点实验室,四川成都610041

出　　处：《中成药》2024年第1期35-40,共6页Chinese Traditional Patent Medicine

基　　金：四川省科技计划项目(2021JDZH0018);四川省科技厅重点研发项目(2022YFS0640);成都市重大科技创新项目(2020-YF08-00042-GX)。

摘　　要：目的研究黄芩茎叶葡萄糖醛酸水解酶(sbslGUS)提取工艺、酶学性质、实际应用。方法在单因素试验基础上,以粒度、加水量、提取时间、提取次数为影响因素,酶活性为评价指标,正交试验优化提取工艺。分析底物(黄芩苷)浓度与酶解速度的关系,计算V_(max)、K_(m),考察pH值、温度、金属离子对酶活性的影响,评价pH稳定性、热稳定性。sbslGUS酶解黄芩苷以制备黄芩素,考察pH值、温度、反应时间、底物初始浓度、加酶量对转化率的影响。结果最优提取工艺为粒度40目,加水量10倍,提取时间15 min,提取次数3次。酶解符合酶促反应动力学,K_(m)为0.0063 mol/L,V_(max)为70.42μmol/h,在pH值6.0、温度45℃、金属离子100 mmol/L Cu2+下酶活性最强,sbslGUS在pH值4.0~7.0、温度4~30℃范围内稳定性良好。最优制备工艺为pH值6.0,温度45℃,反应时间12 h以上,底物初始浓度67.2 mmol/L,加酶量1 mL/0.269 mmol黄芩苷,转化率为97.83%。结论sbslGUS酶解效率高,条件温和,可为获取黄芩素提供简便的制备方法,并且拓展了黄芩茎叶应用途径。AIM To study the extraction process,enzymatic properties and practical application of glucuronic hydrolase in Scutellaria baicalensis stems and leaves(sbsl GUS).METHODS With granularity,water consumption,extraction time and extraction frequency as influencing factors,enzymatic activity as an evaluation index,the extraction process was optimized by orthogonal test on the basis of single factor test.The relationship between substrate(baicalin)concentration and enzymolysis rate,after which V_(max)and K_(m) were calculated,the effects of pH value,temperature and metal ion on enzymatic activity were investigated,pH stability and heat stability were evaluated.sbsl GUS was adotped in the enzymolysis of baicalin to prepare baicalein,then the effects of pH value,temperature,reaction time,initial substrate concentration and enzyme addition on transfer rate were investigated.RESULTS The optimal extraction process was determined to be 40 mesh for granularity,10 times for water consumption,15 min for extraction time,and 3 times for extraction frequency.The enzymolysis accorded with the kinetics of enzymatic reaction,K_(m) was 0.0063 mol/L,V_(max)was 70.42μmol/h,the strongest enzymatic activity was found at the pH value of 6.0,temperature of 45℃and metal ion of 100 mmol/L Cu 2+,sbsl GUS demonstrated good stability at the ranges of 4.0-7.0 for pH value and 4-30℃for temperature.The optimal preparation process was determined to be 6.0 for pH value,45℃for temperature,more than 12 h for reaction time,67.2 mmol/L for initial substrate concentration,and 1 mL/0.269 mmol baicalin for enzyme addition,the transfer rate was 97.83%.CONCLUSION sbsl GUS enzymolysis exhibits high efficiency and mild condition,which can provide a simple preparation method for obtaining baicalein,and expand the application path of Scutellaria baicalensis stems and leaves.

关 键 词：黄芩茎叶 葡萄糖醛酸水解酶 提取工艺 酶学性质 实际应用 

分 类 号：R284[医药卫生—中药学]