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

Elisabeth Hengge, Eva-Maria Steyskal, Alexander Dennig, Manfred Nachtnebel, Harald Matthias Fitzek, Roland Würschum, Bernd Nidetzky*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Article number2400844
JournalSmall Methods
Early online date19 Sept 2024
DOIs
Publication statusE-pub ahead of print - 19 Sept 2024

Keywords

  • bioelectrochemistry and biosensing
  • biointerface engineering
  • nanoporous gold
  • self-assembled monolayer
  • enzyme electrode
  • flavocytochrome P450
  • charge-directed functional immobilization

ASJC Scopus subject areas

  • General Chemistry
  • General Materials Science

Fields of Expertise

  • Advanced Materials Science
  • Human- & Biotechnology

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