Targeted Formation of Biofilms on the Surface of Graphite Electrodes as an Effective Approach to the Development of Biosensors for Early Warning Systems

Author:

Kharkova Anna1ORCID,Perchikov Roman1ORCID,Kurbanalieva Saniyat2,Osina Kristina1,Popova Nadezhda3ORCID,Machulin Andrey4ORCID,Kamanina Olga1,Saverina Evgeniya2,Saltanov Ivan5,Melenkov Sergey5,Butusov Denis6ORCID,Arlyapov Vyacheslav1ORCID

Affiliation:

1. Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia

2. N. D. Zelinsky Institute of Organic Chemistry, 119991 Moscow, Russia

3. Federal State Budgetary Institution of Science Institute of Physical Chemistry and Electrochemistry of the Russian Academy of Sciences, 119071 Moscow, Russia

4. Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences—A Separate Subdivision of the FRC Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences, 142290 Pushchino, Russia

5. Limited Liability Company “INNOBIOSYSTEMS”, 117342 Moscow, Russia

6. Computer-Aided Design Department, Saint Petersburg Electrotechnical University “LETI”, 197022 Saint Petersburg, Russia

Abstract

Biofilms based on bacteria Pseudomonas veronii (Ps. veronii) and Escherichia coli (E. coli) and yeast Saccharomyces cerevisiae (S. cerevisiae) were used for novel biosensor creation for rapid biochemical oxygen demand (BOD) monitoring. Based on the electrochemical measurement results, it was shown that the endogenous mediator in the matrix of E. coli and Ps. veronii biofilms and ferrocene form a two-mediator system that improves electron transport in the system. Biofilms based on Ps. veronii and E. coli had a high biotechnological potential for BOD assessment; bioreceptors based on such biofilms had high sensitivity (the lower limits of detectable BOD5 concentrations were 0.61 (Ps. veronii) and 0.87 (E. coli) mg/dm3) and high efficiency of analysis (a measurement time 5–10 min). The maximum biosensor response based on bacterial biofilms has been observed in the pH range of 6.6–7.2. The greatest protective effect was found for biofilms based on E. coli, which has high long-term stability (151 days for Ps. veronii and 163 days for E. coli). The results of the BOD5 analysis of water samples obtained using the developed biosensors had a high correlation with the results of the standard 5-day method (R2 = 0.9820, number of tested samples is 10 for Ps. veronii, and R2 = 0.9862, number of tested samples is 10 for E. coli). Thus, biosensors based on Ps. veronii biofilms and E. coli biofilms could be a novel analytical system to give early warnings of pollution.

Funder

Ministry of Science and Higher Education of the Russian Federation

Publisher

MDPI AG

Reference47 articles.

1. (2003). ISO Water Quality: Determination of Biochemical Oxygen Demand after n Days (BODn), Part 1: Dilution and Seeding Method with Allylthiourea Addition (Standard No. ISO 5815-1:2003).

2. (2003). ISO Water Quality: Determination of Biochemical Oxygen Demand after n Days (BODn), Part 2: Dilution and Seeding Method with Allylthiourea Addition (Standard No. ISO 5815-1:2003).

3. Arlyapov, V.A., Plekhanova, Y.V., Kamanina, O.A., Nakamura, H., and Reshetilov, A.N. (2022). Microbial Biosensors for Rapid Determination of Biochemical Oxygen Demand: Approaches, Tendencies and Development Prospects. Biosensors, 12.

4. Arlyapov, V.A., Yudina, N.Y., Asulyan, L.D., Kamanina, O.A., Alferov, S.V., Shumsky, A.N., Machulin, A.V., Alferov, V.A., and Reshetilov, A.N. (2020). Registration of BOD Using Paracoccus yeei Bacteria Isolated from Activated Sludge. 3 Biotech, 10.

5. Methods for Assessing Biochemical Oxygen Demand (BOD): A Review;Jouanneau;Water Res.,2014

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