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通过光谱椭圆偏振术和粗粒度模拟研究三油酸甘油酯/水界面和脂肪酶活性。

The triolein/aqueous interface and lipase activity studied by spectroscopic ellipsometry and coarse grained simulations.

机构信息

Physical Chemistry, Department of Chemistry, Lund University, P.O. Box 124, S-221 00 Lund, Sweden; Novozymes A/S, Brudelysvej 26, DK-2880 Bagværd, Denmark.

Novozymes A/S, Brudelysvej 26, DK-2880 Bagværd, Denmark.

出版信息

Chem Phys Lipids. 2018 Mar;211:37-43. doi: 10.1016/j.chemphyslip.2017.10.011. Epub 2017 Nov 10.

Abstract

In spite of the importance of the triglyceride aqueous interface for processes like emulsification, surfactant interactions and lipase activity, relatively little is known about this interface compared to that between alkanes and water. Here, the contact between triolein and water was investigated in terms of water inclusion in the oil phase and orientation of the molecules at the interface. Coarse grained models of triglycerides in contact with water were constructed and correlated with experimental results of the changes in thickness and refractive index, obtained using spectroscopic ellipsometry of spin-coated triolein films. The topography of the layer was revealed by atomic force microscopy. Dry triolein and a triolein sample after equilibration with water were also compared structurally using small-angle X-ray scattering. Additionally, the kinetics of adsorption/activity of three different variants of the Thermomyces lanuginosus lipase (TLL) were investigated. The results show that uptake of water in the triolein phase leads to increase in thickness of the layer. The observed increase of thickness was further enhanced by an active lipase but reduced when an inactive mutant of the enzyme was applied.

摘要

尽管甘油三酯的水界面对于乳化、表面活性剂相互作用和脂肪酶活性等过程非常重要,但与烷烃和水之间的界面相比,人们对该界面的了解相对较少。在这里,研究了三油酸甘油酯与水之间的接触,包括水在油相中的包含以及界面处分子的取向。构建了与实验结果相关联的粗粒度甘油三酯模型,这些实验结果是通过旋涂三油酸甘油酯薄膜的光谱椭圆偏振测量得到的厚度和折射率变化获得的。原子力显微镜揭示了该层的形貌。还使用小角 X 射线散射对干燥的三油酸甘油酯和与水达到平衡后的三油酸甘油酯样品进行了结构比较。此外,还研究了三种不同变体的Thermomyces lanuginosus 脂肪酶(TLL)的吸附/活性动力学。结果表明,三油酸甘油酯相中水分的吸收会导致层厚度的增加。观察到的厚度增加进一步被活性脂肪酶增强,但当使用酶的无活性突变体时,厚度会降低。

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