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为了提高普鲁宁的收率、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、p H、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:柚皮苷浓度13.79 mmol/L、p H=8.0、温度65℃、加酶量18.0 mg/m L、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、收率达到96.87%。连续使用3个批次后,普鲁宁的收率可以达到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,体积分数为40%~60%的异丙醇线性梯度洗脱,从0.36 g柚皮苷反应液中可以分离得到0.22 g普鲁宁。
In order to improve the yield of Prussian, to simplify the process of separation and purification, to obtain a higher purity Prussian, the experiment using naringin crosslinked naringin hydrolysis pruinin, Prunelin concentration is optimized Indicators, the substrate concentration, pH, temperature, amount of enzyme, the reaction time on the preparation of Prussian effect. Orthogonal design was used to optimize the hydrolysis process. The optimum conditions were as follows: naringin concentration 13.79 mmol / L, p H = 8.0, temperature 65 ℃, enzyme dosage 18.0 mg / m L, reaction time 12.0 h, optimization After the preparation of the Prussian concentration of 13.36 mmol / L, the yield reached 96.87%. After using 3 batches continuously, the yield of Prussian can reach 91.29%, and the stability of cross-linked enzyme aggregates is better. The polyamide resin was used for the adsorption of hydrolysates, with a linear gradient of isopropanol with a volume fraction of 40% -60%, and 0.22 g of Prussianin was isolated from 0.36 g of naringin reaction solution.