复合酶法水解大豆浓缩磷脂制备甘油磷脂酰胆碱工艺研究

Preparation of Glycerophosphorylcholine by Complex Enzymatic Hydrolysis of Soybean Concentrated Phospholipids

以大豆浓缩磷脂为原料,复合酶(磷脂酶A1和磷脂酶A2)为催化剂水解制备甘油磷脂酰胆碱。首先建立了甘油磷脂酰胆碱的液相色谱标准曲线,方程为y=8 179.1x-3 492.1(R2=0.998 3);然后测定了大豆浓缩磷脂中磷脂和磷脂酰胆碱的含量,分别为42%和10.4%。通过单因素试验,考察了反应时间、反应温度和加酶比例对复合酶法制备甘油磷脂酰胆碱工艺效果的影响。根据单因素试验结果,选取影响显著的因素,以甘油磷脂酰胆碱转化率为评价指标,采用Box-Benhnken试验设计对工艺条件进行了优化,得到最优工艺条件:反应时间95 min、反应温度41℃、加酶比例(以反应物质量计)1.1%。在此条件下,甘油磷脂酰胆碱转化率为70.65%,与理论预测值基本一致。

Glycerophosphorylcholine was prepared by hydrolysis of soybean concentrated phospholipids using complex enzyme as catalyst in the present study. The HPLC standard curve of glycerophosphorylcholine (GPC) used for the qualitative and quantitative analysis was established,and the equation was y=8 179.1x-3 492.1(R2=0.998 3). The contents of phospholipids and phosphatidylcholine in the raw material were determined,which were 42% and 10.4%,respectively. The effect of reaction time,reaction temperature and enzyme loading ratio on the preparation of GPC by complex enzymatic method were investigated by single factor experiments. Response Surface Methodology(RSM)adopting Box-Benhnken Design(BBD)principle was used to further optimize the preparation processing conditions of GPC on the basis of single factor experiment,with the conversion rate of GPC as evaluation indicator. The optimal conditions for GPC preparation were as follows: reaction time 95 min,reaction temperature 41 ℃ and enzyme loading ratio 1.1%. Under these conditions,the conversion rate of GPC reached to 70.65%,which was nearly consistent with the predicted value.