title: Solid-support immobilization of a "swing" fusion protein for enhanced glucose oxidase catalytic activity.
abstract: 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.
authors: Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Kitakyushu, Fukuoka, 808-0196, Japan; JST ACT-C, Japan.
authors: Yoshiyuki
authors: Y
authors: Takatsuji
authors: Ryota
authors: R
authors: Yamasaki
authors: Atsushi
authors: A
authors: Iwanaga
authors: Michael
authors: M
authors: Lienemann
authors: Markus B
authors: MB
authors: Linder
authors: Tetsuya
authors: T
authors: Haruyama
mesh_terms: D000327:Adsorption; D015374:Biosensing Techniques / Q000379:methods*; D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized / Q000378:metabolism; D005233:Fatty Alcohols; D005656:Fungal Proteins; D005949:Glucose Oxidase / Q000378:metabolism*; D055767:Immobilized Proteins; D058525:Quartz Crystal Microbalance Techniques; D011993:Recombinant Fusion Proteins; D013499:Surface Properties
chemical_list: D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D005233:Fatty Alcohols; D005656:Fungal Proteins; D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; D005949:Glucose Oxidase; C400265:1,2,6-hexanetriol
keywords: Glucose oxidase; Hydorphobin; Self-organized membrane; Surface functionalization