有机溶剂和混合油脂对脂肪酶活性恢复的影响

Effect of Organic Solvents and Mixed Oils on the Recovery of Lipase Activity

本文通过多次催化酯交换方法,研究了最优催化条件下脂肪酶活性和结构的关系,以及低活性脂肪酶的活性再生技术。在催化合成1,3-二油酸-2-棕榈酸甘油三酯的研究中,利用荧光光谱和傅里叶变换红外光谱,分析最优条件下不同使用次数脂肪酶的三级与二级结构,研究脂肪酶活性变化与其二级、三级结构之间的关系。使用异丙醇和混合油脂处理脂肪酶,使低活性脂肪酶的活性得以恢复。研究结果表明:随着脂肪酶活性的降低,脂肪酶荧光强度增强,氨基酸暴露面积逐渐增大,在二级结构中,α-螺旋含量降低,β-折叠含量逐渐增加。通过异丙醇处理,脂肪酶活性恢复到原脂肪酶的75.00%;再经混合油脂处理,脂肪酶活性恢复到原脂肪酶的96.60%,实现了高效催化恢复活性的目的。

In this experiment,through multiple catalytic transesterification methods,the relationship between lipase activity and structure under optimal catalytic conditions and the activity regeneration technology of low-activity lipase are studied.Fluorescence spectroscopy and Fourier transform infrared spectroscopy were used to analyze the tertiary and secondary structures of lipases and the relationship between changes in lipase activity and its secondary tertiary structure.Low-activity lipases were restored by treating lipase with isopropanol and mixed fats.The study results showed that with the decrease of lipase activity,the fluorescence intensity of lipase increased gradually,and the amino acid leakage was more and more.In the secondary structure,theα-helix content decreased and theβ-sheet content increased gradually.Through isopropanol treatment,the lipase activity was restored to 75.00%of the original lipase;through the mixed oil treatment,the lipase activity was restored to 96.60%of the original lipase,realizing the purpose of high-efficiency catalytic restoration.