kk京尼平苷为底物测定β-葡萄糖苷酶活力的方法.pdfVIP

182 2006, Vol. 27, No. 04 梁华正，刘富梁，彭玲西，吴志梅 (东华理工学院生物系，江西 抚州 344000) ：研究以京尼平苷为底物氨基酸为显色剂测定β-葡萄糖苷酶活力的方法。β-葡萄糖苷酶水解京尼平苷的温 度为50℃，pH 为5.0，京尼平苷浓度为0.625mmol/L，水解10min后，立即加入1ml 1mol/L Na2CO3 终止反应， 混匀，再加入体积比为1:1的 0.2mg/ml的精氨酸溶液，沸水浴显色10min，冷却后于 590nm处测光吸收度值。该 方法的检测线性范围为0.05～1U/ml，相关系数为0.9998，检测限为0.02U/ml，精密度为1.5%(n=5)，回收率为99.5%～ 101.1% ，该方法准确度高，结果稳定。 ：β- 葡萄糖苷酶；京尼平苷；精氨酸；活力测定 Method for Determination of β- glucosidase Activity for Geniposide LIANG Hua-zheng，LIU Fu-liang，PENG Ling-xi，WU Zhi-mei (Biology Department, East China Institute of Technology, Fuzhou 344000, China) Abstract ：A method for determination of β- glucosidase activity using geniposide as substrate by amino acid colorimetry was reported. The hydrolysis conditions of geniposide with β- glucosidase and the color reaction conditions, were optimized as: hydrolyzing temperature 50℃, time 10min, pH5.0, color reaction temperature 100℃, color reaction time 10min with the volume ratio of geniposide solvent to Arg solvent 1:1.The method measurement range is 0.05～1U/ml, the linear correlationship 0.9998, the detection limit 0.02U/ml and RSD 1.5%(n=5) with the recovery rate is 99.5% to 101.1%.This method has the advantages of wide measuring range, and the determination of β- glucosidase activity. Key words：beta glucosidase；geniposide；arginine；determination of enzyme activity 中图分类号：Q556.2 文献标识码：A 文章编号：1002-6630(2006)04-0182-04 β-葡萄糖苷酶(EC3.2.1.21)英文名为beta glucosidase， 为无荧光的底物，以此测定；三是以对—硝苯-β- 葡萄 属水解酶类，它能将水果、蔬菜中的风味前体物质β- 糖苷(pNPG)为底物进行酶解，底物水解后释放出来的配 糖苷水解为具有天然风味的香气物质，同时还能协助纤 基对—硝基苯酚可直接在400～9420nm 之间测定。目前 维素酶将纤维素水解为葡萄糖，可用于水解纤维素及生