| 引用本文: | 陈俊,李利君,倪辉,肖安风.重组α-L-鼠李糖苷酶制备普鲁宁[J].中国现代应用药学,2016,33(8):1025-1030. |
| CHEN Jun,LI Lijun,NI Hui,XIAO Anfeng.Transformation of Prunin by Recombinant α-L-rhamnosidase[J].Chin J Mod Appl Pharm(中国现代应用药学),2016,33(8):1025-1030. |
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| 摘要: |
| 目的 利用重组α-L-鼠李糖苷酶，以柚皮苷为底物，通过生物转化法制备普鲁宁。方法 以HPLC检测生物转化反应产物，通过DNS法对酶促反应过程进行动态分析，分析酶促反应动力学，优化普鲁宁制备工艺。结果 重组α-L-鼠李糖苷酶水解柚皮苷反应完全后，可得到反应产物普鲁宁和鼠李糖；生物转化法制备普鲁宁的最适反应温度为60 ℃、pH值为4.0、加酶量为12 U·mL-1、底物浓度为2.0 g·L-1，在此最优工艺条件下，94%的柚皮苷转化为普鲁宁；高浓度的Mn2+、Fe2+能够促进柚皮苷转化生成普鲁宁；重组酶对底物亲和力强，酶促反应的米氏常数Km为1.02 μmol·mL-1，Vmax为0.19 μmol·mL-1·min-1。结论 利用重组α-L-鼠李糖苷酶生物转化法制备普鲁宁，转化率高、重现性好、产物易于分离，可为普鲁宁功能研究、开发利用及改性等提供重要依据。 |
| 关键词: 重组α-L-鼠李糖苷酶 柚皮苷 普鲁宁 生物转化 |
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| 基金项目:福建省教育厅科技项目(JA11160)；福建省科技重点项目(2014Y0081) |
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| Transformation of Prunin by Recombinant α-L-rhamnosidase |
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CHEN Jun1,2, LI Lijun2, NI Hui2, XIAO Anfeng2
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1.College of Chemical Engineering, Huaqiao University, Xiamen 361021, China;2.College of Food and Biological Engineering, Jimei University, Xiamen 361021, China
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| Abstract: |
| OBJECTIVE To investigate the transformation of prunin by recombinant α-L-rhamnosidase. METHODS The final product was identified by HPLC method. The dynamic conversion process was analyzed by DNS method, then the preparation process was optimized. RESULTS The transformation final products were prunin and rhamnose. The optimal conditions for transformation of prunin were 60 ℃, pH 4.0, enzyme dosage 12 U·mL-1, substrate concentration 2.0 g·L-1. Under the optimal condition, 94% naringin could convert to prunin. High concentration of Mn2+ and Fe2+ could significantly accelerate transformation of prunin. The dynamic parameters of Km and Vmax were 1.02 μmol·mL-1 and 0.19 μmol·mL-1·min-1 respectively. CONCLUSION The preparation of prunin by recombinant α-L-rhamnosidase has high conversion rate, good reproducibility, and the products were easy to separate. |
| Key words: recombinant α-L-rhamnosidase naringin pruning transformation |
