固定化柚皮苷酶解转化制备普鲁宁-《集美大学学报（自然科学版）》

文章信息/Info

作者:: 焦超1; 李婧雅1; 李利君1; 2; 杨秋明1; 2; 翁惠芬1; 3; 肖安风1; 2; 3; （1.集美大学食品与生物工程学院，福建厦门 361021；2.福建省食品微生物与酶工程重点实验室，福建厦门 361021；3.福建省海洋功能食品工程技术研究中心，福建厦门 361021）

Author(s):: JIAO Chao1; LI Jingya1; LI Lijun1; 2; YANG Qiuming1; 2; WENG Huifen1; 3; XIAO Anfeng1; 2; 3; (1.College of Food and Biological Engineering,Jimei University,Xiamen 361021,China;2.Key Laboratory of Food Microbiology and Enzyme Engineering of Fujian Province,Xiamen 361021,China;3.Fujian Provincial Engineering Technology Research Center of Marine Functional Food,Xiamen 361021,China)

Keywords:: pruning; substrate immobilization; naringin; enzymatic transformation

摘要:: 选择D101-1大孔吸附树脂固定化反应底物柚皮苷，考察α-L-鼠李糖苷酶水解柚皮苷转化普鲁宁的工艺条件，结果表明，其最适条件为：α-L-鼠李糖苷酶用量30 U/mL，最适酶反应温度60 ℃，pH=4.0，底物质量浓度2.4 g/L，振荡速率80 r/min。在此条件下酶解反应420 min，柚皮苷转化率达到78%。此外，研究发现，高浓度的Mn2+、Fe2+对酶解反应有极强的促进作用。Lineweaver-Burk双倒数拟合曲线的Km=5.12 g/L，Vmax=0.013 g/(L·min)。利用薄层色谱(thin layer chromatography,TLC）和高效液相色谱（high performance liquid chromatography,HPLC）对酶解产物进行分析，并验证得出，反应完全后可得到纯的反应产物普鲁宁。

Abstract:: Naringin immobilized with D101-1 macroporous resin was used as the reaction substrate,the process conditions of α-L-rhamnosidase hydrolysis of naringin to convert pruning were investigated.The conditions for pruning transformation were optimized as α-L-rhamnosidase dosage 30 U / mL,reaction temperature 60 ℃,pH=4.0,substrate concentration 2.4 g/L,shaking rate 80 r/min.Under the optimal conditions,the conversion of naringin reached 78% after enzymatic hydrolysis for 420 min.In addition,it was found that high concentration of Mn2+,Fe2+ could significantly promote the enzymatic reaction.The Km and Vmax of Lineweaver-Burk fitted curve were 5.12 g/L and 0.013 g/(L·min),respectively.The analysis of the hydrolyzate by TLC and HPLC showed that pure pruning could be obtained after the reaction was completed.