Protocol for construction and characterization of direct electron transfer-based enzyme-electrode using gold binding peptide as molecular binder

Hyeryeong Lee; Eun Mi Lee; Stacy Simai Reginald; In Seop Chang

doi:10.1016/j.xpro.2022.101466

Protocol for construction and characterization of direct electron transfer-based enzyme-electrode using gold binding peptide as molecular binder

STAR Protoc. 2022 Jun 14;3(3):101466. doi: 10.1016/j.xpro.2022.101466. eCollection 2022 Sep 16.

Authors

Hyeryeong Lee¹, Eun Mi Lee¹, Stacy Simai Reginald¹, In Seop Chang²

Affiliations

¹ School of Earth Sciences and Environmental Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Gwangju 61005, Republic of Korea.
² School of Earth Sciences and Environmental Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Gwangju 61005, Republic of Korea. Electronic address: ischang@gist.ac.kr.

Abstract

Here, we present a protocol for constructing direct electron transfer (DET)-based enzyme-electrodes using gold-binding peptide (GBP). We describe fusion of four GBPs to flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase gamma-alpha complex (GDHγα), as model oxidoreductase, to generate four GDHγα variants. We then detail the measurements of catalytic and bioelectrochemical properties of these GDHγα variants on electrode together with surface morphology of GDHγα variants immobilized on gold surface. This protocol is useful for construction and validation of enzyme-based electrocatalytic system. For complete details on the use and execution of this protocol, please refer to Lee et al. (2021).

Keywords: Atomic Force Microscopy (AFM); Biophysics; Biotechnology and bioengineering; Material sciences; Microscopy; Molecular Biology; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Electrodes
Electrons
Glucose 1-Dehydrogenase* / genetics
Gold* / chemistry
Peptides / genetics

Substances

Peptides
Gold
Glucose 1-Dehydrogenase