浑源黄芪药材豆腥味与品质关联性探讨——脂肪氧合酶提取及影响酶活因素的研究  

Correlations between Radix Astragali Flavor and Its Quality

作　　者：谢道生[1,2] 武滨 孙海峰[1,2] 郭小青[1] 张丽增[1] 秦雪梅[1,2] 

机构地区：[1]山西大学化学化工学院,太原030006 [2]山西大学中医药现代化研究中心,太原030006 [3]山西省医药集团有限责任公司,太原030012

出　　处：《世界科学技术-中医药现代化》2009年第3期375-381,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：科技部国家科技支撑计划子课题(2006BA106A15-7):山西道地药材黄芪规范化种植及关键技术研究;负责人:武滨;山西优势中药材质量评价与产地适宜性研究(052018);负责人:秦雪梅

摘　　要：目的：研究浑源黄芪药材中豆腥味物质产生的关键生物因子一脂肪氧合酶（Lipoxygenase，LOX）。方法：通过紫外分光光度法测定酶活，在确定黄芪存在LOX活性的基础上，优化黄芪药材LOX提取与测定方法，并考察包括生长年限在内的生物与非生物因素对黄芪LOX活性的影响。结果：浑源黄芪药材中存在LOX活性，最佳提取方法为：液氮研磨黄芪成粉，按照1：7．5的物料比加入pH9．0、O．05mol·L。的硼酸盐缓冲溶液冰水浴浸提1h，低温离心取上清备用：活性测定时酶提取液与底物溶液比例为1：3～1：10（V：V），检测波长为236nm；黄芪LOX在0℃～50℃时较为稳定，70℃时活性急剧下降，Fe^3＋、Fe^2＋、Ca^2＋对该酶有激活作用，Zn^2＋和Mg^2＋则表现为抑制作用，酒石酸也具有较强的抑制作用。浑源基地五年生黄芪的LOX活性最高。结论：浑源产黄芪中存在LOX活性，其活性与温度、金属离子种类和浓度、鳌合剂种类和黄芪生长年限等多种因素有关。Lipoxygenase （LOX） is a key biotic factor in producing the bean-like flavor in Radix Astragali. Authors studied LOX using spectrophotometry and Linolic acid. They developed the needed extraction and detection methods to investigate a range of biotic and abiotic factors, including temperature, metal ion, chelate, and age, and to evaluate LOX activity of Radix Astragali. Results show that LOX exists in the Radix Astragali grown in Hunyuan. The following are the desirable extraction process： Radix Astragali was ground into fine powder using liquid nitrogen, before being soaked for one hour in borate buffer. LOX crude extracts were obtained through centrifugation at 4000 r·min^-1 for 10 at 4℃. LOX crude extracts and substrate solution were mixed in a ratio ranging from 1：10 to 1：3 when LOX activity was detected at a wavelength of 236 nm. Results also suggest that the enzyme was stable at 0℃-50℃ for 15 min, though became inactivated rapidly at 70℃ for 2 min. It can be activated by Fe^3＋, Fe^2＋, and Ca^2＋ , though inhibited by Zn^2＋, Mg^2＋, and tartaric acid. Radix Astragali plants of five years old have the highest LOX activity, suggesting that LOX activity is associated with the growing age. The findings are useful for further studying the mechanism of producing the bean-like flavor in Radix Astragali and authentic sites for growing the plant.

关 键 词：黄芪 豆腥味 脂肪氧合酶 提取 检测 生物与非生物因子 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]