Targeted Formation of Biofilms on the Surface of Graphite Electrodes as an Effective Approach to the Development of Biosensors for Early Warning Systems
Anna Kharkova,
Roman Perchikov,
Saniyat Kurbanalieva,
Kristina Osina,
Nadezhda Popova,
Andrey Machulin,
Olga Kamanina,
Evgeniya Saverina,
Ivan Saltanov,
Sergey Melenkov,
Denis Butusov,
Vyacheslav Arlyapov
Affiliations
Anna Kharkova
Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia
Roman Perchikov
Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia
Saniyat Kurbanalieva
N. D. Zelinsky Institute of Organic Chemistry, 119991 Moscow, Russia
Kristina Osina
Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia
Nadezhda Popova
Federal State Budgetary Institution of Science Institute of Physical Chemistry and Electrochemistry of the Russian Academy of Sciences, 119071 Moscow, Russia
Andrey Machulin
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
Olga Kamanina
Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia
Evgeniya Saverina
N. D. Zelinsky Institute of Organic Chemistry, 119991 Moscow, Russia
Ivan Saltanov
Limited Liability Company “INNOBIOSYSTEMS”, 117342 Moscow, Russia
Sergey Melenkov
Limited Liability Company “INNOBIOSYSTEMS”, 117342 Moscow, Russia
Denis Butusov
Computer-Aided Design Department, Saint Petersburg Electrotechnical University “LETI”, 197022 Saint Petersburg, Russia
Vyacheslav Arlyapov
Federal State Budgetary Educational Institution of Higher Education, Tula State University, 300012 Tula, Russia
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.
