Analytical and Bioanalytical Chemistry

, Volume 381, Issue 8, pp 1558–1567 | Cite as

Monolayers of photosystem II on gold electrodes with enhanced sensor response—effect of porosity and protein layer arrangement

  • J. Maly
  • J. Krejci
  • M. Ilie
  • L. Jakubka
  • J. Masojídek
  • R. Pilloton
  • K. Sameh
  • P. Steffan
  • Z. Stryhal
  • M. Sugiura
Original Paper


Mass transport of the bulk of the analyte to the electrode and through the bioactive layer can be significantly improved by use of the nanoelectrode array and defined arrangement of protein film. This phenomenon has been studied by (i) atomic-force microscopy, (ii) electrochemical measurements of PSII activity, and (iii) digital simulations for an oriented monolayer of histidine-tagged photosystem II (PSII) immobilized on nitrilotriacetic acid (NTA)-modified gold electrodes. The output signal of the electrochemical biosensor is controlled by (i) mass transport from the bioactive layer to electrode and (ii) mass transport between the bulk of the analyte and the electrode. Mass transport through the bioactive layer was electrochemically studied for PSII self-assembled on gold screen-printed electrodes. A densely packed monolayer of PSII has a significant shielding effect toward the diffusion of redox mediator duroquinone (DQ). Mass transport to the planar electrode surface was improved by co-immobilization of bovine-serum albumin (BSA) as spacer biomolecule in the monolayer of PSII. Correlation between the electrochemical properties and surface arrangement of the resulting protein films was clearly observable and confirmed the improved mass-transport properties of structured enzyme monolayers. On the basis of this observation, the application of a bottom-up approach for improvement of electrode performance was proposed and digitally simulated for an infinite array of electrodes ranging in diameter from 50 nm to 5 μm. The nanoelectrode array, with the optimum time window selected for measurements, enables enhancement of mass transport between the bulk of the analyte and the macroelectrode by a factor of up to 50 in comparison with “classical” planar electrodes. Use of a time window enables minimization of crosstalk between individual electrodes in the array. The measurements require methods which suppress the double-layer capacity.


Protein monolayer Photosystem II Nanostructured electrodes Biosensors Mass transport 



Gold working electrode


Bovine serum albumin




Nα,Nα-Bis(carboxymethyl)-l-lysine hydrate








Histidine-tagged photosystem II


Light-emitting diode


Macroelectrode consisting of an array of nanoelectrodes


2-(N-Morpholino)ethanesulfonic acid




Nitrilotriacetic acid


Gold working electrode modified with SAM layer of nitrilotriacetic acid


1-Cyclohexyl-3-(2-morpholinoethyl)carbodiimidemetho-p-toluene sulfonate


Photosystem II


His-PSII immobilized on NTA-AuWE


Self-assembled monolayer


Phosphate buffer



This work was supported by EU project Growth GRD1-2001-41831 “Microprotein”, by project 522/03/0659 of the Grant Agency of the Czech Republic, by project “IBIS” of the Czech Ministry of Industry and Trade, and by the COSMIC Project (ENEA Target Project on Biosensors and Bioelectronics).


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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • J. Maly
    • 1
  • J. Krejci
    • 2
  • M. Ilie
    • 3
  • L. Jakubka
    • 4
  • J. Masojídek
    • 5
  • R. Pilloton
    • 2
  • K. Sameh
    • 4
  • P. Steffan
    • 4
  • Z. Stryhal
    • 1
  • M. Sugiura
    • 6
  1. 1.Department of BiologyUniversity of Jan Evangelista PurkyněÚstí nad LabemCzech Republic
  2. 2.Krejčí EngineeringTišnovCzech Republic
  3. 3.ENEASP061RomaItaly
  4. 4.Department of Microelectronics, CES Centre of Electrochemical Sensors, Joint Research Activity of BVT Technologies and University of TechnologyFEECBrnoCzech Republic
  5. 5.Institute of MicrobiologyAcademy of SciencesTřeboňCzech Republic
  6. 6.Osaka Prefecture UniversityOsakaJapan

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