Microbial mediator biosensors for surface water toxicity determination make it possible to carry out an early assessment of the environmental object’s quality without time-consuming standard procedures based on standard test-organisms, and provide broad opportunities for receptor element modifying depending on the required operational parameters analyzer. Four microorganisms with broad substrate specificity and nine electron acceptors were used to form a receptor system for toxicity assessment. Ferrocene was the most effective mediator according to its high rate constant of interaction with the microorganisms (0.33 ± 0.01 dm3/(g × s) for yeast Saccharomyces cerevisiae). Biosensors were tested on samples containing four heavy metal ions (Cu2+, Zn2+, Pb2+, Cd2+), two phenols (phenol and p-nitrophenol), and three natural water samples. The «ferrocene- Escherichia coli» and «ferrocene-Paracoccus yeei, E. coli association» systems showed good operational stability with a relative standard deviation of 6.9 and 7.3% (14 measurements) and a reproducibility of 7 and 5.2% using copper (II) ions as a reference toxicant. Biosensor analysis with these systems was shown to highly correlate with the results of the standard method using Chlorella algae as a test object. Developed biosensors allow for a valuation of the polluted natural water’s impact on the ecosystem via an assessment of the influence on bacteria and yeast in the receptor system. The systems could be used in toxicological monitoring of natural waters.

The possibility of using the microorganisms Pseudomonas sp. 7p-81, Pseudomonas putida BS394(pBS216), Rhodococcus erythropolis s67, Rhodococcus pyridinivorans 5Ap, Rhodococcus erythropolis X5, Rhodococcus pyridinivorans F5 and Pseudomonas veronii DSM 11331T as the basis of a biosensor for the phenol index to assess water environments was studied. The adaptation of microorganisms to phenol during growth was carried out to increase the selectivity of the analytical system. The most promising microorganisms for biosensor formation were the bacteria P. putida BS394(pBS216). Cells were immobilized in redox-active polymers based on bovine serum albumin modified by ferrocenecarboxaldehyde and based on a composite with a carbon nanotube to increase sensitivity. The rate constants of the interaction of the redox-active polymer and the composite based on it with the biomaterial were 193.8 and 502.8 dm3/(g·s) respectively. For the biosensor created using hydrogel bovine serum albumin-ferrocene-carbon nanotubes, the lower limit of the determined phenol concentrations was 1 × 10−3 mg/dm3, the sensitivity coefficient was (5.8 ± 0.2)∙10−3 μA·dm3/mg, Michaelis constant KM = 230 mg/dm3, the maximum rate of the enzymatic reaction Rmax = 217 µA and the long-term stability of the bioanalyzer was 11 days. As a result of approbation, it was found that the urban water phenol content differed insignificantly, measured by creating a biosensor and using the standard photometric method.