交联柚苷酶聚集体水解柚皮苷制备普鲁宁

Hydrolysis of Naringin to Prunin by Cross-linked Naringinase Aggregates

摘要:

为了提高普鲁宁的得率、简化分离提纯的工艺、得到较高纯度的普鲁宁，本实验采用交联柚苷酶聚集体水解柚皮苷制备普鲁宁，以普鲁宁浓度为优化指标，研究了底物浓度、pH、温度、加酶量、反应时间对制备普鲁宁的影响。利用正交设计对水解过程进行优化，确定最佳工艺条件为：底物浓度13.79 mmol/L、pH值8.0、温度65 °C、加酶量18.0 mg/mL、反应时间12.0 h，优化后制备得到的普鲁宁浓度为13.36 mmol/L、得率达到96.87 %。连续使用3个批次，普鲁宁的得率可以达到 91.29 %，交联酶聚集体的稳定性较好。采用聚酰胺树脂对水解产物进行吸附，40~60%异丙醇线性梯度洗脱，可以有效地将普鲁宁分离，为普鲁宁制备提供了一种新的方法。

Abstract:

In order to improve the prunin yield, simplify the process of separation and purification and obtain the high purity production, the conversion of naringin to prunin by cross-linked naringinase aggregate was investigated. The effects of substrate concentration, pH, temperature, enzyme dosage and the reaction time on the preparation of prunin by cross-linked naringinase aggregates were optimized, and the prunin concentration was took as the optimization index. The orthogonal design was used to optimize the catalytic processes, and the optimum conditions for producing the prunin were as follow: substrate concentration 13.79 mmol/L, pH 8.0, temperature 65 °C, enzyme dosage 18.0 mg/mL, reaction time 12.0 h. Under the above conditions, the production of prunin was about 13.36 mmol/L, and the yield of prunin was 96.87 %. The cross-linked aggregates was stable, it could successively convert for three cycles, and the yield of prunin could reach 91.29 %. Prunin can be effectively separated from the hydrolysis products by using the polyamide resin adsorption and 40% to 60 % isopropanol linear gradient elution .It provided a new approach for preparing the prunin.