Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Kitakyushu, Fukuoka, 808-0196, Japan; JST ACT-C, Japan.
Colloids Surf B Biointerfaces. 2013 Dec 1;112:186-91. doi: 10.1016/j.colsurfb.2013.07.051. Epub 2013 Aug 2.
The strategic surface immobilization of a protein can add new functionality to a solid substrate; however, protein activity, e.g., enzymatic activity, can be drastically decreased on immobilization onto a solid surface. The concept of a designed and optimized "molecular interface" is herein introduced in order to address this problem. In this study, molecular interface was designed and constructed with the aim of attaining high enzymatic activity of a solid-surface-immobilized a using the hydrophobin HFBI protein in conjunction with a fusion protein of HFBI attached to glucose oxidase (GOx). The ability of HFBI to form a self-organized membrane on a solid surface in addition to its adhesion properties makes it an ideal candidate for immobilization. The developed fusion protein was also able to form an organized membrane, and its structure and immobilized state on a solid surface were investigated using QCM-D measurements. This method of immobilization showed retention of high enzymatic activity and the ability to control the density of the immobilized enzyme. In this study, we demonstrated the importance of the design and construction of molecular interface for numerous purposes. This method of protein immobilization could be utilized for preparation of high throughput products requiring structurally ordered molecular interfaces, in addition to many other applications.
蛋白质的策略性表面固定化可以为固体基质添加新的功能；然而，蛋白质的活性，例如酶活性，在固定到固体表面上时可能会大大降低。为了解决这个问题，本文引入了设计和优化“分子界面”的概念。在这项研究中，设计并构建了分子界面，目的是通过使用疏水蛋白 HFBI 与附着在葡萄糖氧化酶 (GOx) 上的 HFBI 融合蛋白，实现固定化表面上 a 的高酶活性。HFBI 除了具有粘附特性外，还能够在固体表面上形成自组织膜，这使其成为固定化的理想候选物。开发的融合蛋白也能够形成有组织的膜，并使用 QCM-D 测量研究了其在固体表面上的结构和固定状态。这种固定化方法显示出保留高酶活性和控制固定化酶密度的能力。在这项研究中，我们证明了设计和构建分子界面对于许多目的的重要性。这种蛋白质固定化方法可用于制备需要结构有序的分子界面的高通量产品，以及许多其他应用。
