固定化柚皮苷酶解转化制备普鲁宁

Preparation of Pruning by Immobilized Naringin

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Mn^(2+)、Fe^(2+)对酶解反应有极强的促进作用。Lineweaver-Burk双倒数拟合曲线的K_m=5.12 g/L,V_(max)=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。

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 Mn 2+,Fe 2+ could significantly promote the enzymatic reaction.The K m and V max 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.