猪胰脂肪酶固定化载体的优化

Research on the Optimization of Immobilized Vector of PPL

[目的]考察不同固定化载体与酶量的比例对固定化效果的影响以及缓冲溶液pH值对固定化效果的影响。[方法]以大孔树脂、硅藻土和活性炭的不同加入量作为因素,同时以缓冲溶液的不同pH值作为另外一个因素来考察对固定效果的影响。[结果]通过采用经典的橄榄油乳化法对脂肪酶水解活力进行测定,将得出的结果进行综合性比较。最终试验结果表明,利用大孔树脂的物理吸附法获得了较高的固定化酶水解活性收率。在固定化温度为30℃条件下,以大孔树脂作为载体时固定化猪胰脂肪酶效果最佳,此时缓冲溶液K2HPO4-KH2PO4的pH值为6.4,大孔树脂与猪胰脂肪酶的质量比为2.5∶1.0。如对反应温度、反应时间以及用酶量进行适当的控制,将可以得到活性更高的固定化脂肪酶。[结论]研究得出,固定化载体的选择以及缓冲溶液的pH值都是影响固定化效果的重要因素。

[ Objective] The effect of the different immobilized carriers, proportions of enzyme and pH value of buffer on hanaobilization was studied. [ Method] The effect of both the different amounts of macroporous resins, diatomaceous earth and activated carbon, and the different pH values of buffer on immobilization was tested. [ Results] The vitality of lipase hydrolysis was determined with the classic method of olive oil emulsion and the results were com- prehensively compared. Final test results showed that the high recovery ratio of the hydrolysis activity of immobilized enzyme was high in the physical adsorption method of macrooporous resin. At imnobilized temperature of 30 ℃, the effect of ro, acroporous resin as a carrier on the immobilization of PPL was the best,at that tnoraent, the pH value of buffer K2HPO4-KH2PO4 solution was 6.4 and the ratio of macroporons resin to PPL was 2.5：1.0.The higher activity of immobilized lipase could be produced if the reaction telrrperature,reaction time and the enzyme amotmt were appropriately controlled. [Conclusion] The vector and the pH value of buffer solution were important factors affecting immobilization of PPL.