Choi, Okkyoung Kim, Byung Chun An, Ji-Hye Min, Kyoungseon Kim, Yong Hwan Um, Youngsoon Oh, Min-Kyu Sang, Byoung-In 2024-01-20T16:04:26Z 2024-01-20T16:04:26Z 2021-09-05 2011-10 0141-0229 https://pubs.kist.re.kr/handle/201004/129931 We constructed a fusion protein (GOx-R5) consisting of R5 (a polypeptide component of silaffin) and glucose oxidase (GOx) that was expressed in Pichia pastoris. Silaffin proteins are responsible for the formation of a silica-based cell matrix of diatoms, and synthetic variants of the R5 protein can perform silicification in vitro [1]. GOx secreted by P. pastoris was self-immobilized (biosilicification) in a pH 5 citric buffer using 0.1 M tetramethoxysilane as a silica source. This self-entrapment property of GOx-R5 was used to immobilize GOx on a graphite rod electrode. An electric cell designed as a biosensor was prepared to monitor the glucose concentrations. The electric cell consisted of an Ag/AgCl reference electrode, a platinum counter electrode, and a working electrode modified with poly(neutral red) (PNR)/GOx/Nafion. Glucose oxidase was immobilized by fused protein on poly(neutral red) and covered by Nafion to protect diffusion to the solution. The morphology of the resulting composite PNR/GOx/Nafion material was analyzed by scanning electron microscopy (SEM). This amperometric transducer was characterized electrochemically using cyclic voltammetry and amperometry in the presence of glucose. An image produced by scanning electron microscopy supported the formation of a PNR/GOx complex and the current was increased to 1.58 mu A cm(-1) by adding 1 mM glucose at an applied potential of -0.5 V. The current was detected by way of PNR-reduced hydrogen peroxide, a product of the glucose oxidation by GOx. The detection limit was 0.67 mM (S/N = 3). The biosensor containing the graphite rod/PNR/GOx/Nafion detected glucose at various concentrations in mixed samples, which contained interfering molecules. In this study, we report the first expression of R5 fused to glucose oxidase in eukaryotic cells and demonstrate an application of self-entrapped GOx to a glucose biosensor. Crown Copyright (C) 2011 Published by Elsevier Inc. All rights reserved. English ELSEVIER SCIENCE INC A biosensor based on the self-entrapment of glucose oxidase within biomimetic silica nanoparticles induced by a fusion enzyme Article 10.1016/j.enzmictec.2011.07.005 1 ENZYME AND MICROBIAL TECHNOLOGY, v.49, no.5, pp.441 - 445 ENZYME AND MICROBIAL TECHNOLOGY 49 5 441 445 N scie scopus 000297396000002 2-s2.0-81155159804 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article CARBON-FILM ELECTRODES SOL-GEL MATERIALS IMMOBILIZATION COMPOSITES BIOSILICA NANOTUBE ROUTE Biosilicification Enzyme immobilization Fused enzyme Silaffin Glucose oxidase Biosensor