亳菊花中β-葡萄糖苷酶的提取和酶学性质研究  

Extraction and Enzymatic Properties of β-Glucosidase in Chrysanthemum morifolium

作　　者：左亚锋 张伟[1,2,3] 吴德玲 俞年军[1,2] 金传山[1,2] 刘劲松[1,2,3] 朱月健[4] 闫攀[1] 

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]安徽省现代中药重点实验室,安徽合肥230012 [3]安徽省中医药科学院药物化学研究所,安徽合肥230012 [4]安徽济人药业有限公司,安徽亳州236000

出　　处：《安徽中医药大学学报》2015年第5期83-86,共4页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金青年项目(81303229);安徽省自然科学基金项目(1408085MH20);安徽省科技攻关项目(1301042100)

摘　　要：目的测定亳菊花中β-葡萄糖苷酶的活性，研究亳菊花中β葡萄糖苷酶的基本酶学性质。方法采用对硝基苯酚-β-D葡萄糖苷（4-nitrophenyl-β-D-glucopyranoside，pNPG）法考察亳菊花中β-葡萄糖苷酶活性测定的最佳条件及其酶学性质。结果当0．2mL酶液、20mmol／L pNPG0．2mL、0．6mL pH值为4．0的柠檬酸-柠檬酸三钠缓冲液在温度为50℃的水浴中反应45min时，酶活性最高。在此条件下对亳菊花的粗酶液中8I葡萄糖苷酶的基本酶学性质进行初步研究，结果表明该酶在30～50℃时，随着温度升高，酶活力有上升趋势，其热稳定性较好；pH值在3．2～5．4之间时，相对酶活性均在80％以上；酶在4℃下的保存稳定性较好。结论本研究所测定的亳菊花中β葡萄糖苷酶活性的最佳条件及酶学性质可为亳菊花炮制原理研究奠定基础。Objective To determine the activity of β-glucosidase in Chrysanthemum morifolium, and to study the fundamental enzymatic properties of β-glucosidase in Chrysanthemum morifolium. Methods The β-nitrophenyl- D-glucoside （pNPG） method was used to measure the optimal condition for determination of the activity of β-glucosidase in Chrysanthemum morifolium and its enzymatic properties. Results The β-glucosidase in Chrysanthemum morifolium had the highest activity when the mixture of 0.2 mL enzyme solution, 0.2 mL pPNG （20 mmol/L）, and 0.6 mL citric acid-trisodium citrate buffer （pH 4.0） was incubated for 45 rain in a 50 ℃ water bath. The fundamental enzymatic properties of β-glucosidase in the crude enzyme solution of Chrysanthemum morifolium were preliminarily studied under the above condition. The results showed that the enzymatic activity increased as the temperature increased from 30 ℃ to 50 ℃, suggesting a high thermal stability of the enzyme. The relative activity of the enzyme kept higher than 80 % at pH 3.2-5.4. The enzyme was quite stable during storage at 4 ℃. Conclusion The optimal condition for the activity of β-glucosidase in Chrysanthemum. morifolium and its enzymatic properties determined in this study provide a basis for the processing principle of Chrysanthemum morifolium.

关 键 词：亳菊花 对硝基苯酚-β—D葡萄糖苷法 β-葡萄糖苷酶活性 

分 类 号：R284.2[医药卫生—中药学]