Electrodes for enzyme immobilization

  • Projects


  • eBioTech - Enzyme, Electrode and Reactor Design for Enzymatic Cascade Reactions


(DFG project, 01.05.2024 - 30.04.2027)

In this joint project, funded by the DFG and running in the frame of the DFG Priority Programm (SPP) 2240 eBioTech, we develop enzyme (Prof. Oliver Einsle, University of Freiburg), electrode (Prof. Anna Fischer, University of Freiburg) and reactor systems (Prof. Ulf-Peter Apfel, Ruhr Universiät Bochum) for enzymatic cascade reactions and in particular for the bioelectrocatalytic nitrate ammonification. 

DFG Priority Program (SPP) 2240 eBiotech



  • CarboBioElec - Bioelectrocatalytic conversion of hydrocarbons with enzyme and whole-cell electrode biohybrids

(BMBF project, 01.10.2020 - 31.03.2024)

In the project CarboBioElec, funded by the BMBF, we develop advanced 3D porous electrode systems for the immobilization and direct wiring of electroactive enzymes and whole cell systems for bioelectrocatalytic applications.  The enzymatic and whole cell systems are thereby developed by the group of Prof. Dr. Boll (University of Freiburg), while single enzyme structure determination by cryo EM is performed in the group of Prof. Wendler (University Potsdam). Electrode development and immobilization strategies are thereby developed in our group.



  • Nanostructured Transparent Conductive Electrodes – Advanced Material Design For Controlled Enzyme Immobilization


Nanostructured transparent conductive thin film electrodes are promising platforms for the further development of enzymatic bio-electronic hybrid devices, such as enzymatic fuel cells or biosensors.

High surface area materials by design, mesoporous transparent conductive thin film electrodes allow the quantitative immobilization of high amounts of enzyme / proteins only if the pore structure (in terms of pore size and pore connectivity) is designed in agreement with the enzyme / protein size.

As such the aim of our project is to developp high-surface area transparent conductive thin film electrodes with tailored porosity for the quantitative immobilization of redox active enzymes as high performance electrocatalysts.

Moreover by taking advantage of the transparency of these electrodes fundamental insights into relevant surface immobilization processes involving the material-enzyme interface can be gained by spectroscopic techniques.

As such combined electrochemical and spectroscopic investigation of selected enzymes on model as well as high-surface area transparent conductive electrodes allow to unravel material parameters controlling enzyme binding as well as orientation for direct electrochemistry.

We also employ our electrode platforms for the immobilization of molecular electrocatalysts.



This project is funded by the DFG (Cluster of Excellence "Unifying Concepts in Catalysis" UniCat EXC 314-2)



1) Enzymes on ATO (antimony doped tin oxide) electrodes:

Frasca, S.; Molero Milan, A.; Guiet, A.; Goebel, C.; Pérez-Caballero, F.; Stiba, K.; Leimkühler, S.; Fischer*, A.; Wollenberger*, U.
Bioelectrocatalysis at Mesoporous Antimony Doped Tin Oxide Electrodes—Electrochemical Characterization and Direct Enzyme Communication.
Electrochim. Acta 2013, 110, 172–180.


Full-size image (102 K)


Mesostructure and crystallinity of the pl-ATO (a, c) and ATO-F127 films (b, d): SEM images (cross section and top view) of the planar ATO (a) and the ATO-F127 (b) thin films; HRTEM image with the corresponding SAED patterns inset for pl-ATO (c) and ATO-F127 (d); (e) wide-angle XRD of the as-synthesized ATO NPs (A) and of the ATO NPs calcined at 450 °C (B). (doi:10.1016/j.electacta.2013.03.144)


2) Hydrogenases on nanostructured gold electrodes - Immobilization Study by SEIRA spectroelectrochemistry

Heidary, N.; Utesch, T.; Zerball, M.; Horch, M.; Millo, D.; Fritsch, J.; Lenz, O.; von Klitzing, R.; Hildebrandt, P.; Fischer*, A.; Mroginski*, M. A.; Zebger*, I;
Orientation-Controlled Electrocatalytic Efficiency of an Adsorbed Oxygen-Tolerant Hydrogenase.
PLoS One 2015, 10, 1–13.



Fig 1. Top: SEIRA spectra of the Strep-tagged Re MBH, immobilized on a nanostructured Au surface coated with a self-assembled monolayer (SAM) of 6-amino-1-hexanethiol. Spectra are shown for (A), the amide mode region and, (B), the CO and CN stretching region of the active site. A structural depiction of the active site in the oxidized Nir-B state is shown in the inset. Bottom: (C) Non-contact-mode AFM topographic mapping of the SAM-modified Au surface after completed MBH immobilization. The dashed vertical line indicates the course of the height profiles shown in (D) for immobilized Re MBH (orange line) and prior to immobilization (black line; see Fig B in S1 Appendix). The grey double-headed arrow indicates the space of a single MBH molecule.


3) Hydrogenases on surface modified gold electrodes - Interface building by diazonium salt electrografting, enzyme immobilization, protein film voltammetry and SEIRA spectroelectrochemistry

Harris, T. G. A. A.; Heidary, N.; Kozuch, J.; Frielingsdorf, S.; Lenz, O.; Mroginski, M.; Hildebrandt, P.; Zebger, I.*; Fischer, A.*
In Situ Spectroelectrochemical Studies into the Formation and Stability of Robust Diazonium-Derived Interfaces on Gold Electrodes for the Immobilization of an Oxygen-Tolerant Hydrogenase.
ACS Appl. Mater. Interfaces 2018, 10 (27), 23380–23391.






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