Biosurfactant-Producing Bacteria Isolated from a Microbial Consortium Previously Subjected to Adaptive Laboratory Evolution in Oily Sludge

Microbial bioprospecting in contaminated environments is a promising strategy for identifying biosurfactant-producing bacteria; however, translating environmentally adapted strains into predictable cultivation processes remains challenging. In this study, a microbial consortium subjected to long-term evolutionary laboratory adaptation in oily sludge was investigated to evaluate strain-specific phenotypic responses related to biosurfactant production. Phylogenetic analysis based on 16S rDNA sequencing identified three taxonomically distant isolates: Faucicola sp. strain BS5C, Pseudomonas sp. strain BS16B, and Enterobacter sp. BS14MR. Biosurfactant production was evaluated using a sequential Design of Experiments (DOE) approach, including fractional factorial and central composite rotatable designs, with the emulsification index (E24) used as a semi-quantitative response variable. Initial screening revealed a statistically significant negative effect (p < 0.10) of high dextrose concentrations for all isolates. Strain-specific differences in model adequacy were observed, with a statistically adequate quadratic model obtained for Pseudomonas sp. BS16B (R2 = 0.8658, p = 0.0225), whereas the other isolates showed significant lack of fit (p < 0.05). ATR-FTIR analysis revealed spectral profiles consistent with lipopeptide-like compounds. Overall, these results indicate that isolates derived from the same long-term adapted system may differ substantially in process predictability, suggesting that productivity-based screening alone may be insufficient for selecting robust strains.