Electrochemically Induced Nanoscale Stirring Boosts Functional Immobilization of Flavocytochrome P450 BM3 on Nanoporous Gold Electrodes

Abstract: Enzyme‐modified electrodes are core components of electrochemical biosensors for diagnostic and environmental analytics and have promising applications in bioelectrocatalysis. Despite huge research efforts spanning decades, design of enzyme electrodes for superior performance remains challenging. Nanoporous gold (npAu) represents advanced electrode material due to high surface‐to‐volume ratio, tunable porosity, and intrinsic redox activity, yet its coupling with enzyme catalysis is complex. Here, the study reports a flexible‐modular approach to modify npAu with functional enzymes by combined material and protein engineering and use a tailored assortment of surface and in‐solution methodologies for characterization. Self‐assembled monolayer (SAM) of mercaptoethanesulfonic acid primes the npAu surface for electrostatic adsorption of the target enzyme (flavocytochrome P450 BM3; CYT102A1) that is specially equipped with a cationic protein module for directed binding to anionic surfaces. Modulation of the SAM surface charge is achieved by electrochemistry. The electrode‐adsorbed enzyme retains well the activity (33%) and selectivity (complete) from in‐solution. Electrochemically triggered nanoscale stirring in the internal porous network of npAu‐SAM enhances speed (2.5‐fold) and yield (3.0‐fold) of the enzyme immobilization. Biocatalytic reaction is fueled from the electrode via regeneration of its reduced coenzyme (NADPH). Collectively, the study presents a modular design of npAu‐based enzyme electrode that can support flexible bioelectrochemistry applications.

Standort
Deutsche Nationalbibliothek Frankfurt am Main
Umfang
Online-Ressource
Sprache
Englisch

Erschienen in
Electrochemically Induced Nanoscale Stirring Boosts Functional Immobilization of Flavocytochrome P450 BM3 on Nanoporous Gold Electrodes ; day:19 ; month:09 ; year:2024 ; extent:13
Small Methods ; (19.09.2024) (gesamt 13)

Urheber
Hengge, Elisabeth
Steyskal, Eva‐Maria
Dennig, Alexander
Nachtnebel, Manfred
Fitzek, Harald
Würschum, Roland
Nidetzky, Bernd

DOI
10.1002/smtd.202400844
URN
urn:nbn:de:101:1-2409201412540.593753988615
Rechteinformation
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Letzte Aktualisierung
15.08.2025, 07:32 MESZ

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