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Biosurfactants/bioemulsifiers (BSs/BEs) can be defined as surface-active biomolecules produced by microorganisms with a broad range of applications. In recent years, due to their unique properties like biodegradability, specificity, low toxicity, and relative ease of preparation, these biomolecules have attracted wide interest as an eco-friendly alternative for several industrial sectors, escalating global microbial BS/BE market growth. Recently, Gordonia alkanivorans strain 1B, a bacterium with significant biotechnological potential, well known for its biodesulfurizing properties, carotenoid production, and broad catabolic range, was described as a BS/BE producer. This study focuses on the characterization of the properties of the lipoglycopeptide BSs/BEs produced by strain 1B, henceforth referred to as gordofactin, to better understand its potential and future applications. Strain 1B was cultivated in a chemostat using fructose as a carbon source to stimulate gordofactin production, and different purification methods were tested. The most purified sample, designated as extracted gordofactin, after lyophilization, presented a specific emulsifying activity of 9.5 U/mg and a critical micelle concentration of 13.5 mg/L. FT-IR analysis revealed the presence of basic hydroxyl, carboxyl, ether, amine/amide functional groups, and alkyl aliphatic chains, which is consistent with its lipoglycopeptide nature (60% lipids, 19.6% carbohydrates, and 9% proteins). Gordofactin displayed remarkable stability and retained emulsifying activity across a broad range of temperatures (30 °C to 80 °C) and pH (pH 3-12). Moreover, a significant tolerance of gordofactin emulsifying activity (EA) to a wide range of NaCl concentrations (1 to 100 g/L) was demonstrated. Although with a great loss of EA in the presence of NaCl concentrations above 2.5%, gordofactin could still tolerate up to 100 g/L NaCl, maintaining about 16% of its initial EA for up to 7 days. Furthermore, gordofactin exhibited growth inhibition against both Gram-positive and Gram-negative bacteria, and it demonstrated concentration-dependent free radical scavenging activity for 2,2-diphenyl-1-picrylhydrazyl (IC 50 ≈ 1471 mg/L). These promising features emphasize the robustness and potential of gordofactin as an eco-friendly BS/BE alternative to conventional surfactants/emulsifiers for different industrial applications.

Carotenoids are high added-value products primarily known for their intense coloration and high antioxidant activity. They can be extracted from a variety of natural sources, such as plants, animals, microalgae, yeasts, and bacteria. Gordonia alkanivorans strain 1B is a bacterium recognized as a hyper-pigment producer. However, due to its adaptations to its natural habitat, hydrocarbon-contaminated soils, strain 1B is resistant to different organic solvents, making carotenoid extraction through conventional methods more laborious and inefficient. Ionic liquids (ILs) have been abundantly shown to increase carotenoid extraction in plants, microalgae, and yeast; however, there is limited information regarding bacterial carotenoid extraction, especially for the Gordonia genus. Therefore, the main goal of this study was to evaluate the potential of ILs to mediate bacterial carotenoid extraction and develop a method to achieve higher yields with fewer pre-processing steps. In this context, an initial screening was performed with biomass of strain 1B and nineteen different ILs in various conditions, revealing that tributyl(ethyl)phosphonium diethyl phosphate (IL#18), combined with ethyl acetate (EAc) as a co-solvent, presented the highest level of carotenoid extraction. Afterward, to better understand the process and optimize the extraction results, two experimental designs were performed, varying the amounts of IL#18 and EAc used. These allowed the establishment of 50 µL of IL#18 with 1125 µL of EAc, for 400 µL of biomass (cell suspension with about 36 g/L), as the ideal conditions to achieve maximal carotenoid extraction. Compared to the conventional extraction method using DMSO, this novel procedure eliminates the need for biomass drying, reduces extraction temperatures from 50 °C to 22 ± 2 °C, and increases carotenoid extraction by 264%, allowing a near-complete recovery of carotenoids contained in the biomass. These results highlight the great potential of ILs for bacterial carotenoid extraction, increasing the process efficiency, while potentially reducing energy consumption, related costs, and emissions.