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As one of the main non-covalent relations in microbiological-based systems, cell surface hydrophobicity (CSH) can be observed as a relevant parameter for biodegradation capability and suggested bacterial behaviour and biofilm formation during a bioremediation process. On the other hand, the role of ureolytic bacteria in bioremediation has subsequently led to the examination of this bacterial type in different engineering fields. In order to optimize the operating parameters of microbial adhesion to hydrocarbons test (MATH) for ureolytic bacteria, Box–Behnken experimental design was conducted for five ureolytic bacteria isolated from soils, as well as for the reference strain Sporosarcina pasteurii DSM 33. The optimization was completed with and without the essential substrate for the targeted metabolic reaction, with the aim to compare differences in bacterial hydrophobicity. A vortex time of 2 min, a hydrocarbon volume of 0.5 mL, and a phase separation time of 15 min are recommended as MATH operating parameters for all tested ureolytic bacteria. Although all bacteria are hydrophobic, lower CSH values in the presence of urea were observed for the same bacterium, which could be explained by the interaction of urea with the organic phase of the separation system, as well as a rapid ureolysis process that also occurs during the application of ureolytic bacteria in biotechnology systems.
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