Izvestiya of Saratov University.


ISSN 1817-3020 (Print)
ISSN 2542-193X (Online)

For citation:

Kozlowski A. V., Stetsyura S. V. Features of photo-stimulated adsorption of enzymes on semiconductor substrate. Izvestiya of Saratov University. Physics , 2023, vol. 23, iss. 4, pp. 316-327. DOI: 10.18500/1817-3020-2023-23-4-316-327, EDN: JMIHFG

This is an open access article distributed under the terms of Creative Commons Attribution 4.0 International License (CC-BY 4.0).
Published online: 
Full text:
(downloads: 80)
Article type: 

Features of photo-stimulated adsorption of enzymes on semiconductor substrate

Kozlowski Alexander V., Saratov State University
Stetsyura Svetlana Viktorovna, Saratov State University

Background and Objectives: Features of photostimulated adsorption of enzymes on a semiconductor substrate, leading to different changes in the sensitivity to glucose and hydrogen peroxide, were studied using the enzymes glucose oxidase and horseradish peroxidase as an example. Materials and Methods: Enzyme molecules were deposited on n-Si and p-Si substrates by photostimulated layer-by-layer adsorption from solution. Glucose oxidase and horseradish peroxidase were used as enzymes. The resulting structures were mounted in an electrochemical cell to measure the capacitance-voltage characteristics of the electrolyte-insulator-semiconductor contact, which were then used to determine the sensitivity of the sensor structures to glucose and hydrogen peroxide. Results: The results were analyzed taking into account photoelectronic processes in n-Si and p-Si semiconductor substrates. An increase in the sensitivity to the analyte from the use of photostimulated adsorption has been found for the structures obtained on the basis of n-Si, regardless of the type of immobilized enzyme. But for glucose oxidase molecules, the effect of photostimulation reaches 200%, and for horseradish peroxidase molecules it does not exceed 30%. The effect of photostimulated adsorption is explained by the charge exchange of surface electronic states at the Si/SiO2 interface upon illumination and the formation of induced dipoles that combine the charge of the enzyme molecule and the opposite charge of the Si/SiO2 interface after the illumination is turned off. Conclusion: The conducted studies can be applied in the development of a capacitive biosensor operating on the field effect, since taking into account the change in the charge state of the immobilized enzyme and the surface of the semiconductor signal converter makes it possible in some cases to significantly increase the sensitivity of the biosensor.

The work was supported by the Russian Science Foundation (project No. 22-22-00194, https://rscf.ru/en/project/22-22-00194/).
  1. Malyar I. V., Gorin D. A., Santer S., Stetsyura S. V. Photocontrolled Adsorption of Polyelectrolyte Molecules on a Silicon Substrate. Langmuir, 2013, vol. 29, iss. 52, pp. 16058–16065. https://doi.org/10.1021/la403838n
  2. Malyar I. V., Santer S., Stetsyura S. V. The Effect of Illumination on the Parameters of the Polymer Layer Deposited from Solution onto a Semiconductor Substrate. Technical Physics Letters, 2013, vol. 39, iss. 7, pp. 656–659. https://doi.org/10.1134/S1063785013070183
  3. Stetsyura S. V., Kozlowski A. V., Malyar I. V. The influence of a silicon substrate conductivity type on the efficiency of photostimulated polyelectrolyte adsorption. Technical Physics Letters, 2017, vol. 43, iss. 3, pp. 376–379. https://doi.org/10.1134/S1063785017040290
  4. Kozlowski A. V., Stetsyura S. V. Formation feature of organic polyelectrolyte layer on illuminated semiconductor substrate. Izvestya of Saratov University. Physics, 2022, vol. 22, iss. 3, рр. 254–265 (in Russian). https://doi.org/10.18500/1817-3020-2022-22-3-254-265
  5. Dobrynin A. V., Deshkovski A., Rubinstein M. Adsorption of Polyelectrolytes at an Oppositely Charged Surface. Phys. Rev. Lett., 2000, vol. 84, iss. 14, pp. 3101–3104. https://doi.org/10.1103/PhysRevLett.84.3101
  6. Dobrynin A. V., Rubinstein M. Theory of polyelectrolytes in solutions and at surfaces. Prog. Polym. Sci., 2005, vol. 30, iss. 11, pp. 1049–1118. https://doi.org/10.1016/j.progpolymsci.2005.07.006
  7. Stetsyura S. V., Kozlowski A. V. The influence of photoelectron processes in a semiconductor substrate on the adsorption of polycationic and polyanionic molecules. Technical Physics Letters, 2017, vol. 43, iss. 3, pp. 285–288. https://doi.org/10.1134/S1063785017030233
  8. Kozlowski A. V., Stetsyura S. V. Kinetics of photostimulated adsorption of enzyme molecules onto n- and p-type silicon. IOP Conference Series : Materials Science and Engineering, 2019, vol. 699, article no. 012022 (4 p.). https://doi.org/10.1088/1757-899X/699/1/012022
  9. Grigorenko V. G., Andreeva I. P., Rubtsova M. Yu., Egorov A. M. Recombinant horseradish peroxidase: Production and analytical applications, Biochemistry (Moscow), 2015, vol. 80, no. 4, pp. 408–416. https://doi.org/10.1134/S0006297915040033
  10. Portaccio M., Lepore M. Determination of Different Saccharides Concentration by Means of a MultienzymesAmperometric Biosensor. Journal of Sensors, 2017, vol. 2017, article no. 7498945 (8 p.). https://doi.org/10.1155/2017/7498945
  11. Kaygorodov K. L., Smirnova M. A., Tarabanko V. E. Synthesis of Divanillin in the Presence of Water Extract and Juice of Horseradish Root. Journal of Siberian Federal University. Сhemistry, 2020, vol. 13, no. 4, рp. 525–533 (in Russian). https://doi.org/10.17516/1998-2836-0195
  12. Harris J. M., Reyes C., Lopez G. P. Common Causes of Glucose Oxidase Instability in Vivo Biosensing: A Brief Review. Journal of Diabetes Science and Technology, 2013, vol. 7, no. 4, article no. 1030 (8 р.). https://doi.org/10.1177/193229681300700428
  13. Hecht H. J., Kalisz H. M., Hendle J., Schmid R. D., Schomburg D. Crystal structure of glucose oxidase from Aspergillus niger refined at 2.3 A resolution. J. Mol. Biol., 1993, vol. 229, iss. 1, pp. 153–172. https://doi.org/10.1006/jmbi.1993.1015
  14. Xie Y., Li Z., Zhou J. Hamiltonian replica exchange simulations of glucose oxidase adsorption on charged surfaces. Physical Chemistry Chemical Physics, 2018, vol. 20, iss. 21, рp. 14587–14596. https://doi.org/10.1039/C8CP00530C
  15. Maslennikova A. A., Kozlowski A. V., Santer S., Stetsyura S. V. The influence of illumination and ionic strength of a solution on the formation of biosensor structure based on a silicon substrate and glucose oxidase molecules. Journal of Physics: Conference Series, 2019, vol. 1400, article no. 077052 (6 p.) https://doi.org/10.1088/1742-6596/1400/7/077052
  16. Tan S., Gu D., Liu H., Liu Q. Detection of a single enzyme molecule based on a solid-state nanopore sensor. Nanotechnology, 2016, vol. 27. no. 15, article no. 1555021 (11 р.) https://doi.org/10.1088/0957-4484/27/15/155502
  17. Ahirwal G. K., Mitra C. K. Direct Electrochemistry of Horseradish Peroxidase-Gold Nanoparticles Conjugate. Sensors, 2009, vol. 9, iss. 2, рp. 881–894. https://doi.org/10.3390/s90200881
  18. Nandini S., Nalini S., Manjunatha R., Shanmugam S., Melo J. S., Suresh G. S. Electrochemical biosensor for the selective determination of hydrogen peroxide based on the co-deposition of palladium, horseradish peroxidase on functionalized-graphene modified graphite electrode as composite. Journal of Electroanalytical Chemistry, 2013, vol. 689, pp. 233–242. https://doi.org/10.1016/j.jelechem.2012.11.004
  19. Krainer F. W., Glieder A. An updated view on horseradish peroxidases: Recombinant production and biotechnological applications. Applied Microbiology and Biotechnology, 2015, vol. 99, рp. 1611–1625. https://doi.org/10.1007/s00253-014-6346-7
  20. Ferapontova E., Domínguez E. Adsorption of differently charged forms of horseradish peroxidase on metal electrodes of different nature: Effect of surface charges. Bioelectrochemistry, 2002, vol. 55, iss. 1–2, рp. 127–130. https://doi.org/10.1016/S1567-5394(01)00155-4
  21. Rennke H. G., Venkatachalam M. A. Chemical modification of horseradish peroxidase. Preparation and characterization of tracer enzymes with different isoelectric points. Journal of Histochemistry & Cytochemistry, 1979, vol. 27, iss. 10, рp. 1352–1353. https://doi.org/10.1177/27.10.41873
  22. Cloarec J. P., Chevalier C., Genest J., Beauvais J., Chamas H., Chevolot Y., Baron T., Souifi A. pH driven addressing of silicon nanowires onto Si3N4/SiO2 micropatterned surfaces. Nanotechnology, 2016, vol. 27, article no. 295602 (10 р.). https://doi.org/10.1088/0957-4484/27/29/295602
  23. Movillia J., Huskens J. Functionalized Polyelectrolytes for Bioengineered Interfaces and Biosensing Applications. Organic Materials, 2020, vol. 2, iss. 2, pp. 78–107. https://doi.org/10.1055/s-0040-1708494
  24. Poghossian A., Abouzar M. H., Amberger F., Mayer D., Han Y., Ingebrandt S., Offenhausser A., Schoning M. J. Field-effect sensors with charged macromolecules: Characterisation by. Biosensors and Bioelectronics, 2007, vol. 22, iss. 9–10, pp. 2100–2107. https://doi.org/10.1016/j.bios.2006.09.014
  25. Garyfallou G. Z., de Smet L. C. P. M., Sudhölter E. J. R. The effect of the Type of doping on the electrical characteristics of electrolyte–oxide–silicon sensors: pH sensing and polyelectrolyte adsorption. Sensors and Actuators B: Chemical, 2012, vol. 168, pp. 207–213. https://doi.org/10.1016/j.snb.2012.04.010
  26. Aué J., de Hosson J. T. Influence of atomic force microscope tip-sample interaction on the study of scaling behavior. Applied Physics Letters, 1997, vol. 71, iss. 10, рp. 1347–1349. https://doi.org/10.1063/1.120415
  27. Makky A., Viel P., Chen S. W., Berthelot T., Pellequer J., Polesel-Maris J. Piezoelectric tuning fork probe for atomic force microscopy imaging and specific recognition force spectroscopy of an enzyme and its ligand. J. Mol. Recognit., 2013, vol. 26, iss. 11, pp. 521–531. https://doi.org/10.1002/jmr.2294