Crude Biosurfactant Research Articles

Valorization of oil refinery by-products: production of sophorolipids utilizing fatty acid distillates and their potential antibacterial, anti-biofilm, and antifungal activities.

Starmerella bombicola is a native yeast strain producing sophorolipids as secondary metabolites. This study explores the production, characterization, and biological activities of sophorolipids and investigates the antimicrobial, anti-biofilm, and antifungal properties of sophorolipids produced from oil refinery wastes by the yeast Starmerella bombicola. The present work demonstrated that S. bombicola MTCC 1910 when grown in oil refinery wastes namely palm fatty acid distillates and soy fatty acid distillates enhanced the rate of sophorolipids production drastically in comparison to vegetable oil, sunflower oil used as hydrophobic feedstock. Sophorolipid yields were 18.14, 37.21, and 46.1g/L with sunflower oil, palm, and soy fatty acid distillates respectively. The crude biosurfactants were characterized using TLC, FTIR, and HPLC revealing to be acetylated sophorolipids containing both the acidic and lactonic isomeric forms. The surface lowering and emulsifying properties of the sophorolipids from refinery wastes were significantly higher than the sunflower oil-derived sophorolipids. Also, all the sophorolipids exhibited strong antibacterial properties (minimum inhibitory concentrations were between 50 and 200µgmL-1) against Salmonella typhimurium, Bacillus cereus, and Staphylococcus epidermidis and were validated with morphological analysis by Scanning electron microscopy. All the sophorolipids were potent biofilm inhibitors and eradicators (minimum biofilm inhibitory and eradication concentrations were between12.5to1000µgmL-1) for all the tested organisms. Furthermore, antifungal activities were also found to exhibit about 16-56% inhibition at 1mgmL-1 for fungal mycelial growth. Therefore, this endeavour of sophorolipids production using palm and soy fatty acid distillates not only opens up a window for the bioconversion of industrial wastes into productive biosurfactants but also concludes that sophorolipids from oil refinery wastes are potent antimicrobial, anti-biofilm, and antifungal agents, highlighting their potential in biotechnological and medical applications.

Candida parapsilosis CMGB-YT Biosurfactant for Treatment of Heavy Metal- and Microbial-Contaminated Wastewater

During the last few decades, water pollution has become a growing concern at international level. To date, only a few Candida parapsilosis strains were successfully used in environmental remediation. In the present article, the strain C. parapsilosis CMGB-YT was studied for its ability to assimilate hydrophobic substrates and to produce biosurfactants with antimicrobial activity and positive effects on heavy metal removal from contaminated wastewaters. The strain C. parapsilosis CMGB-YT was grown on yeast peptone (YP) media with 1% n-decane, n-dodecane, n-tetradecane, n-hexadecane, as well as commercial sunflower and olive oils. The production of the biosurfactant was evaluated using the emulsification index (E24%). The surface properties and emulsifying stability of the biosurfactant were determined. The effect of the biosurfactant on the cell growth of two strains of Rhodotorula mucilaginosa and on their removal capacity of lead (0.032 g/L) and cadmium (0.030 g/L) ions from synthetic wastewater were also studied. The antimicrobial potential of 20 mg/mL and 40 mg/mL biosurfactant was established in the presence of pathogenic Candida krusei strains. C. parapsilosis CMGB-YT assimilated n-hexadecane with good rates over 216 h and produced an anionic glycolipidic biosurfactant with stable E24% towards long-chain carbon compounds at different temperatures, with an alkaline pH and high salinity (10% NaCl). The biosurfactant reduced the surface tension to 53.58 ± 0.42 mN/m, while the critical micellar concentration (CMC) was reached at 4.2% biosurfactant. The crude biosurfactant (5%) enhanced R. mucilaginosa growth in heavy metal-contaminated wastewater, increased chemical oxygen demand (COD) removal of up to 80%, and improved Cd²⁺ removal by 10%. Additionally, the concentrated biosurfactant effectively prevented Candida krusei biofilm formation. In conclusion, the biosurfactant produced by C. parapsilosis CMGB-YT demonstrates promising potential for the efficient treatment of wastewater contaminated with heavy metals and microbial pathogens.

Screening of sustainable biosurfactant produced by indigenous bacteria isolated from Egyptian oil fields for microbial enhanced oil recovery

Microbial enhanced oil recovery (MEOR) stands out as an environmentally and economically significant approach utilizing microorganisms and their bio-products to reduce residual oil, and hence improve oil recovery. Due to the pollution that synthetic surfactants entail and their very expensive costs, there is a growing necessity for clean, eco-friendly alternatives. This research focuses on exploring the potential of producing biosurfactants by indigenous bacteria isolated from Egyptian Oil Fields. Five crude oil and formation water samples were collected from four Egyptian Oil Fields located in the Western Desert (WD). Nine bacterial strains were isolated and screened for biosurfactant production using different assays, followed by the extraction and purification of crude biosurfactants. The most promising biosurfactant-producing strains that showed their ability to produce effective biosurfactant according to biosurfactant screening assays were selected for further studies and identified through 16S rRNA gene sequencing. Furthermore, core-flooding micromodel experiments were conducted to evaluate the effectiveness of these extracted biosurfactants compared to a synthetic surfactant, Sodium Dodecyl Sulfate (SDS), in improving oil recovery. Results showed that the most promising biosurfactant-producing strains that exhibited notable performance and ability to produce effective biosurfactants were namely A-LB, C-LB2, and B-YM2. Therefore, they were selected for further studies and identified through 16S rRNA gene sequencing. 16S rRNA gene sequencing results showed that namely A-LB, C-LB2, and B-YM2 were identified as Bacillus massiliigabonensis, Pseudomonas nitritolerans, and Aninetobacter seohaensis, respectively. Finally, core flooding results showed that the biosurfactant produced by Bacillus massiliigabonensis demonstrated significant performance by recovering 69.96% of additional oil, whereas 68.11% and 63.34% of additional oil were recovered using biosurfactants produced by Pseudomonas nitritolerans, and Aninetobacter seohaensis, respectively. It was also noticed that the biosurfactant produced by Bacillus massiliigabonensis demonstrated significant potential in improving oil recovery compared to the synthetic surfactant (61.78%), it outperformed 8.18% more additional oil. These findings validate the effectiveness of the produced biosurfactants over synthetic surfactants emphasizing their potential as sustainable and cost-effective alternatives for enhanced oil recovery in Egyptian oil fields.

Enhancement of oil biodegradation by using the biosurfactant produced from local Bacillus subtilis isolate

Background: Oil contamination poses a significant threat to the global environment and has attracted widespread attention in recent years. Given its significance, further exploration of biodegradation options for this contamination has never been more crucial than it is today. Therefore, the objective of the study was to improve the biodegradation of oil by utilizing local bacterial isolates alongside a biosurfactant produced by a Bacillus subtilis isolate. Twenty-two bacterial isolates were collected from four samples of hydrocarbon-contaminated soil. These isolates underwent screening to assess their effectiveness in degrading crude oil using two distinct methods.Methods: Soli samples were collected from two contaminated sites with oil pollutants in Baghdad city. Biodegradation ability was tested on liquid Bushnell Haas medium (BH), pH 7 supplemented with 1% of crude oil and then screened primarily using the 2,6-dichlorophenol indophenol method to determine the ability of isolates to degrade the crude oil. All isolates were identified morphologically and biochemically. Biosurfactants was extracted from Bacillus subtilis previously isolated.Results: Isolate SCS1 has appeared the highest ability reaching 52.6% compared with other isolates, also, the results of secondary screening confirmed that isolate SCS1 has given the best biodegradation reaching 53.8%. The isolate SCS1 was identified as Pseudomonas aeruginosa, this isolate was used to study the effect of biosurfactant on crude oil biodegradation. Results exhibited a high biodegradation efficiency reaching 85.1% in culture broths supplemented with 100 mg/100. It was noted that the use of 150 and 200 mg of biosurfactant led to a decrease in the biodegradation of crude oil.Conclusion: The use of biosurfactant led to an increase in the degradation rate by up to 61.7% of crude oil by Pseudomonas aeruginosa when using 25-100 mg of crude biosurfactant that produced by local isolate Bacillus subtilis.Keywords: Biodegradation; Biosurfactant; Bacillus subtilis; Pseudomonas aeruginosa

Isolation and Characterization of Glycolipid Biosurfactant Produced by Marine Bacterium Cobetia marina Strain F1 and Investigation of Antimicrobial and Anti-Biofilm Activity

The marine environment is a rich source of microorganisms producing bioactive compounds, like biosurfactant-producing bacteria that exhibit unique characteristics and functionalities. In this study, we examined glycolipid biosurfactants produced by bacteria that live commensally with marine organisms. We isolated a biosurfactant-producing strain identified as Cobetia marina strain F1, which displayed high hemolytic activity (27 mm), oil spreading ability (4 mm), emulsification index (40%), and decreasing surface tension to 31.3 (mN·m−1). Fourier transform infrared (FT-IR) spectroscopy revealed the glycolipid composition of the biosurfactant. Furthermore, elemental analysis utilizing CHNS and energy-dispersive X-ray spectroscopy (EDS) confirmed the biosurfactant contained carbon, hydrogen, nitrogen, sulfur, chlorine, potassium, oxygen, and additional elements. The critical micelle concentration (CMC) of the crude biosurfactant was determined to be 350 mg·L−1, at which concentration, a decrease in surface tension was observed when the biosurfactant was dissolved in distilled water. Given the presence of impurities in the biosurfactant composition, this observed CMC is considered acceptable. Furthermore, the biosurfactant exhibited significant antimicrobial activity against both Gram-positive and Gram-negative bacterial strains, with the largest zone of inhibition (ZOI) of 27 mm against Pseudomonas aeruginosa. This demonstrates the potential of the biosurfactant to serve as an alternative to novel antibiotic agents. The biosurfactant exhibited considerable inhibition of biofilm formation, disruption of preformed biofilms, and reduced enzymatic activity in bacterial cells following treatment. Moreover, the combination of the biosurfactant and F1 bacterial strain enhanced the degradation of crude oil by 86%, indicating its potential application in environmental remediation. These findings highlight the importance of investigating commensal strains capable to produce biosurfactants for applications in hydrocarbon remediation, overcoming antibiotic resistance, and biofilm disruption.

Yarrowia lipolytica CMGB32 Biosurfactants Produced Using n-Hexadecane: Developing Strategies for Environmental Remediation

The yeast Yarrowia lipolytica degrades petroleum compounds, including alkanes, via the monoterminal oxidation pathway, the hydrophobic carbon substrate assimilation is mediated by biosurfactants, and extracellular amphiphilic molecules are produced by the yeast cell. This study focuses on the ability of the strain Y. lipolytica CMGB32 to degrade n-hexadecane by producing biosurfactants with high potential for bioremediation. The hydrocarbon-degrading potential of the yeast strain was observed via a 2,6-dichlorophenolindophenol (DCPIP) test in Bushnell–Hass medium with 1% n-hexadecane, and cell hydrophobicity was expressed as microbial adhesion to hydrocarbons (MATH). Biosurfactant production on yeast peptone (YP) with 1% n-hexadecane was estimated after 72 h using the emulsification index (E24%) against toluene. Crude biosurfactant (cell-free broth) stability tests were performed at different temperatures (4 °C, 70 °C) and NaCl concentrations (2–10%). The effects of a biosurfactant on synthetic wastewater remediation comprised the growth curves (OD measurements) of natural heavy metal degrader Rhodotorula mucilaginosa, determination of nutrients (spectrophotometrically), physico-chemical parameters, and removal capacity of lead and cadmium ions (via inductively coupled plasma mass spectrometry—ICP-MS). The antimicrobial and anti-adherence activities of 20 mg/mL and 40 mg/mL of the biosurfactant against pathogenic Candida krusei strains involved growth observations and the crystal violet microtiter method. The DCPIP decolorization occurred after six days, corresponding to the maximum growth phase of the Y. lipolytica culture. After 72 h, the cells presented high hydrophobicity (82.61% MATH) and stable biosurfactant production (E24% 47%). The crude biosurfactant (5%) increased the growth of R. mucilaginosa strains cultivated on synthetic wastewater cultures contaminated with Pb2+ and Cd2+, increased the conductivity and COD (86%) of the samples, and determined Pb2+ (66%) and Cd2+ (42%) ions reduction. The concentrated biosurfactant inhibited C. krusei growth (70%) and biofilm adherence. In conclusion, Y. lipolytica CMGB32 shows important potential for development of biosurfactant-based technologies for the remediation of heavy-metal- and emerging pathogen-contaminated wastewaters.

Polyurethane degradation by extracellular urethanase producing bacterial isolate Moraxella catarrhalis strain BMPPS3

Plastic waste has become a global issue and a threat to the ecosystem. The present study isolated polyurethane (PU) degrading bacterial species from soil dumped with plastic wastes. Four bacterial isolates, RS1, RS6, RS9 and RS13 were obtained and their ability to degrade PU in a synthetic medium with PU as a sole source of carbon was assessed individually. After thirty days of incubation, the highest PU weight loss of 67.36 ± 0.32% was recorded in the medium containing RS13 isolate. The results of FTIR revealed the occurrence of carbonyl peaks. The putative isolate RS13 confirmed with the genus Moraxella according to 16S rRNA gene sequencing and the isolate was specified as Moraxella catarrhalis strain BMPPS3. The restriction analysis of Moraxella catarrhalis strain BMPPS3 revealed that the GCAT content to 51% and 49% correspondingly. Moraxella catarrhalis strain BMPPS3 was able to colonize on PU surface and form a biofilm as revealed by SEM investigation. Fatty acids and alkanes were found to be the degradation products by GC-MS analysis. The presence of these metabolites facilitated the growth of strain RS13 and suggested that ester hydrolysis products had been mineralized into CO2 and H2O. Extracellular biosurfactant synthesis has also been found in Moraxella catarrhalis strain BMPPS13 inoculated with synthetic media and mineral salt media containing PU and glucose as carbon sources, respectively with a significant level of cell-surface hydrophobicity (32%). The production and activity of extracellular esterase showed consistent increase from day 1–15 which peaked (1.029 mM/min/mg) on day 24 significantly at P < 0.001. Crude biosurfactants were lipopeptide-based, according to the characteristic investigation. According to this study findings, Moraxella catarrhalis produces biosurfactants of the esterase, urethanase and lipase (lipopeptide) types when carbon source PU is present.

Chemical and biological evaluation of biosurfactant fractions from Wickerhamomyces anomalus CCMA 0358.

Biosurfactants (BS) are becoming a solution for today's world since they are considered a reasonable and eco-friendly option for use in products that require surfactants. This study aimed to evaluate the antibacterial activity of purified fractions containing biosurfactants produced by the yeast Wickerhamomyces anomalus CCMA 0358 using waste cooking oil (WCO) as substrate. Mixed fractions were separated and characterized by TLC, MPLC, GC-MS, LC-OMS, LC-SQMS, FTIR, 1H, 13C, DEPT 135, COSY, HSQC, and HMBC. The results confirmed the presence of palmitic acid and oleic acid fatty acids, derived from the core biosurfactant structure; however, the core could not be identified. The crude biosurfactant and its purified fractions were evaluated against pathogenic bacteria, and the purified fractions of the biosurfactant are more efficient at inhibitory and bactericidal activities than the crude biosurfactant. To the best of our knowledge, this is the first study that evaluated the antimicrobial activity of purified fractions of biosurfactants produced by the species Wickerhamomyces anomalus. Therefore, the purification of biosurfactants can emerge as an interesting alternative to increase the bioactivity of the compounds and ensure greater efficiency and biotechnological employability. KEY POINTS: • Successful production of a biosurfactant using a renewed carbon source. • Evaluation of the antimicrobial activity of purified fractions of BS. • Separated fractions of the BS are more efficient against bacteria than the crude BS.

Optimization and purification of bioproducts from Bacillus velezensis PhCL fermentation and their potential on industrial application and bioremediation

Bioproduction is considered a promising alternative way of obtaining useful and green chemicals. However, the downstream process of biomolecules has been one of the major difficulties in upscaling the application of bioproducts due to the high purification cost. Acid precipitation is the most common method for purifying biosurfactants from the fermentation broth with high purity. However, the use of strong acids and organic solvents in solvent extraction has limited its application. Hence, in this study, a new strain of Bacillus velezensis PhCL was isolated from phenolic waste, and its production of amylase had been optimized via response surface methodology. After that, amylase and biosurfactant were purified by sequential ammonium sulfate precipitation and the result suggested that even though the purified crude biosurfactant had a lower purification fold compared to the acid precipitation, the yield was higher and both enzymes and biosurfactant also could be recovered for lowering the purification cost. Moreover, the purified amylase and crude biosurfactant were characterized and the results suggested that the purified crude biosurfactant would have a higher emulsion activity and petroleum hydrocarbon removal rate compared to traditional surfactants. This study provided another approach for purifying bioactive compounds including enzymes and biosurfactants from the same fermentation broth and further explored the potential of the crude purified biosurfactant in the bioremediation of polycyclic aromatic hydrocarbons and petroleum hydrocarbons.

Development of Nanotechnology-Based Drug Delivery Systems for Controlling Clinical Multidrug-Resistant Staphylococcus aureus and Escherichia coli Associated with Aerobic Vaginitis.

The growing prevalence of resistance to antibiotics potentially makes Escherichia coli and Staphylococcus aureus serious pathogens, necessitating the development of new antimicrobial agents. We extracted crude biosurfactants from a potential probiotic Bacillus spp. to control pathogenic bacteria associated with aerobic vaginal infection. Using nanotechnology formulations, we developed nanoemulsions based on biosurfactants at different concentrations (1% and 3.33%). The results showed that these nanoemulsions were stable, with a weighted index of 0.3, and demonstrated broad-spectrum antibacterial activity against Escherichia coli and Staphylococcus aureus, with MICs ranging between 1.25 and 4 mg/mL. Additionally, the nanoemulsions exhibited interesting antibiofilm effects. All strains became more sensitive to the antibiotics to which they were resistant because of various biosurfactant formulations combined with antibiotics. Lower concentrations of BNE1% and 3.33% were still more efficient than the crude biosurfactants. Our findings demonstrated that the biosurfactant had a strong antibiofilm effect against all tested pathogens. This antibacterial effect can be explained by their ability to alter cell physiology such as cell hydrophobicity and membrane disintegration. Thus, we can conclude that the use of nanotechnology formulations has improved this effect, and the nanoemulsions developed in this study can be used as a potential anti-infectious therapy against multidrug-resistant bacterial strains of clinical origin.

Lichenysin-like Polypeptide Production by Bacillus licheniformis B3-15 and Its Antiadhesive and Antibiofilm Properties.

We report the ability of the crude biosurfactant (BS B3-15), produced by the marine, thermotolerant Bacillus licheniformis B3-15, to hinder the adhesion and biofilm formation of Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213 to polystyrene and human cells. First, we attempted to increase the BS yield, optimizing the culture conditions, and evaluated the surface-active properties of cell-free supernatants. Under phosphate deprivation (0.06 mM) and 5% saccharose, the yield of BS (1.5 g/L) increased by 37%, which could be explained by the earlier (12 h) increase in lchAA expression compared to the non-optimized condition (48 h). Without exerting any anti-bacterial activity, BS (300 µg/mL) prevented the adhesion of P. aeruginosa and S. aureus to polystyrene (47% and 36%, respectively) and disrupted the preformed biofilms, being more efficient against S. aureus (47%) than P. aeruginosa (26%). When added to human cells, the BS reduced the adhesion of P. aeruginosa and S. aureus (10× and 100,000× CFU/mL, respectively) without altering the epithelial cells' viability. As it is not cytotoxic, BS B3-15 could be useful to prevent or remove bacterial biofilms in several medical and non-medical applications.

Production of a New Biosurfactant by a New Yeast Species Isolated from Prunus mume Sieb. et Zucc.

Biosurfactants reduce surface and interfacial tension due to their amphiphilic properties and are an eco-friendly alternative for chemical surfactants. In this study, a new yeast strain JAF-11 that produces a biosurfactant was selected using drop collapse method, and the properties of the extracts were investigated. The nucleotide sequences of the strain were compared with closely related strains and identified based on the D1/D2 domain of the large subunit ribosomal DNA (LSU) and internal transcribed spacer (ITS) regions. Neodothiora populina CPC 39399T, the closest species with strain JAF-11, showed a sequence similarity of 97.75% for LSU and 94.27% for ITS, respectively. The result suggests that the strain JAF-11 represents a distinct species that cannot be assigned to any existing genus or species in the family Dothideaceae. Strain JAF-11 produced a biosurfactant reducing the surface tension of water from 72 mN/m to 34.5 mN/m on the sixth day of culture and the result of measuring the critical micelle concentration (CMC) by extracting the crude biosurfactant was found to be 24 mg/l. The molecular weight 502 of the purified biosurfactant was confirmed by measuring the fast atom bombardment mass spectrum. The chemical structure was analyzed by measuring 1H nuclear magnetic resonance (NMR), 13C NMR, and two-dimensional NMRs of the compound. The molecular formula was C26H46O9, and it was composed of one octanoyl group and two hexanoyl groups to myo-inositol moiety. The new biosurfactant is the first report of a compound produced by a new yeast strain, JAF-11.

A biosurfactant-producing novel bacterial strain isolated from vermicompost having multiple plant growth-promoting traits.

The nutrient-rich vermicompost which is used as manure for the growth and development of plants is rich in microbial flora. These microbes protect the plants against several infectious pathogenic microbes. As certain microbes are known to produce biosurfactants as metabolites, an investigation was carried out to isolate biosurfactant-producing bacterial strains from vermicompost with the efficient antifungal property. From the study, it was revealed that biosurfactant-producing bacterial strains are present in the vermicompost. A total of nine bacterial strains were isolated from the vermicompost. Among them, one most efficient biosurfactant-producing bacterial strains with antifungal properties have been screened. After molecular characterization of the isolated strain, it was revealed that the bacterial strain is Bacillus licheniformis strain SCV1. The strain produces 3.4 ± 0.1 g/L of crude biosurfactant, which when column purified yields 3.1 ± 0.1 g/L of biosurfactant. The biosurfactant exhibited excellent emulsifying activity (E24 ) of 96.56% against crude oil. The produced biosurfactant was identified as a lipopeptide consisting of a mixer of surfactin and iturin. Furthermore, the biosurfactant exhibited significant antifungal activity against a wide range of phytopathogens, showing 76.3% inhibition against Sclerotinia sclerotiorum, 53% inhibition against Colletotrichum gloeosporioides, 51% against Fusarium verticillioides, and 36% against Corynespora cassicolla. Along with antifungal activities, the stain was found to exhibit multiple plant growth-promoting traits. This study, thus indicates that vermicompost might contain biosurfactant-producing microbes which can render protection to the plant against various phytopathogens by the production of biosurfactants and can also stimulate plant growth.

Bioremediation of Wastewater Using Yeast Strains: An Assessment of Contaminant Removal Efficiency.

The main goal of wastewater treatment is to significantly reduce organic compounds, micronutrients (nitrogen and phosphorus) and heavy metals and other contaminants (pathogens, pharmaceuticals and industrial chemicals). In this work, the efficiency of removing different contaminants (COD, NO3-, NO2-, NH4+, PO43-, SO42-, Pb2+, Cd2+) from synthetic wastewater was tested using five different yeast strains: Kluyveromyces marxianus CMGBP16 (P1), Saccharomyces cerevisiae S228C (P2), Saccharomyces cerevisiae CM6B70 (P3), Saccharomyces cerevisiae CMGB234 (P4) and Pichia anomala CMGB88 (P5). The results showed a removal efficiency of up to 70% of COD, 97% of nitrate, 80% of nitrite, 93% of phosphate and 70% of sulfate ions for synthetic wastewater contaminated with Pb2+ (43 mg/L) and Cd2+ ions (39 mg/L). In contrast, the results showed an increase in ammonium ions, especially in the presence of Pb2+ ions. The yeast strains showed a high capacity to reduce Pb2+ (up to 96%) and Cd2+ (up to 40%) ions compared to the initial concentrations. In presence of a crude biosurfactant, the removal efficiency increased up to 99% for Pb2+ and 56% for Cd2+ simultaneously with an increase in yeast biomass of up to 11 times. The results, which were obtained in the absence of aeration and in neutral pH conditions, proved a high potential for practical applications in the biotreatment of the wastewater and the recovery of Pb and Cd ions, with a high benefit-cost ratio.

Efficacious use of potential biosurfactant producing plant growth promoting rhizobacteria to combat petrol toxicity in Zea mays L. plants.

Soil pollution caused by petroleum hydrocarbons is a serious threat for human life, as it affects the groundwater, cause economical losses after decreasing the agricultural production, and create other ecological issues. Here, we are reporting the isolation and screening of rhizosphere bacteria possessing biosurfactant producing potential and capable of enhancing plant growth under petrol stress as well as possessing. Efficient biosurfactant producers having plant growth promoting traits were characterized morphologically, physiologically, and phylogenetically. These selected isolates were identified as Bacillus albus S2i, Paraclostridium benzoelyticum Pb4, and Proteus mirabilis Th1 based on 16S rRNA sequence analysis. These bacteria possessed plant growth promoting attributes as well as exhibited positive activity toward the assays based on hydrophobicity, lipase activity, surface activity, and hydrocarbon degradation that indicated the production of biosurfactants. Fourier transform infrared spectroscopy of crude biosurfactants extracted from bacterial strains revealed that the biosurfactants from Pb4 and Th1 might belong to glycolipid or glycolipopeptide class whereas the biosurfactants from S2i could be from phospholipid class. Scanning electron micrographs exhibited group of exopolymer matrices interconnecting the cells forming a complex network of mass, while energy dispersive X-ray analysis has shown elemental composition of biosurfactants with dominance of nitrogen, carbon, oxygen, and phosphorous. Further, these strains were then used to ascertain their effect on the growth and biochemical parameters including stress metabolites and antioxidant enzymology of Zea mays L. plants grown under petrol (gasoline) stress. Significant increments in all studied parameters were observed in comparison with control treatments that might be due to petrol degradation by bacteria and also by secreting growth stimulating substances released by these bacteria in soil ecosystem. To the best of our knowledge, this is the first report in which Pb4 and Th1 were studied as surfactant producing PGPR and further their role as biofertilizer for the significant improvement in phytochemical constituents of maize plants grown under petrol stress was assessed.

Use of biosurfactants produced by Bacillus subtilis H1 and Pseudomonas aeruginosa PAO1 as a disinfectant and plant growth stimulation

Sustainable agriculture involves the maximum use of the resource potential of the earth, with the constant renewal of the fertility of the ecosystem. One method of transition to sustainable agriculture is the use of biological control agents, which include biosurfactants. We assessed the possibility of using crude biosurfactants obtained from Bacillus subtilis H1 and Pseudomonas aeruginosa PAO1 as dressing agents for untreated wheat seeds and the presence of seed germination stimulating properties. It was shown that crude rhamnolipids obtained from Pseudomonas aeruginosa PAO1 inhibited the area occupied by fungal mycelia at a concentration of 500 mg/l. Surfactin, derived from Bacillus subtilis H1, inhibited fungal growth at a concentration of 100 mg/L. Stimulation of germination of wheat seeds by 1.9 and 2 times was determined by rhamnolipid at a concentration of 500 mg/l and surfactin at a concentration of 100 mg/l for 168 hours of germination. The use of biocontrol agents is a promising method that can improve product quality while reducing the negative impact on the environment.

Experimental investigation for treating ibuprofen and triclosan by biosurfactant from domestic wastewater

The presence of emerging pollutants of pharmaceutical products and personal care products (PPCPs) in the aquatic environment overspreads the threat on living beings. Bioremediation is a promising option for treating wastewater. In the present study, an experimental investigation was carried out to produce a biosurfactant by Pseudomonas aeruginosa (MTCC 1688) for the removal of Ibuprofen (IBU) and Triclosan (TCS) from domestic wastewater. It was performed in three stages. Firstly, the production and optimization of biosurfactant was carried out to arrive at the best combination of crude sunflower oil, sucrose and ammonium bicarbonate (10%: 5.5 g/L: 1 g/L) to yield effective biosurfactant production (crude biosurfactant) and further extended to achieve critical micelle concentration (CMC) formation by dilution (biosurfactant at 10.5%). The stability of the biosurfactant was also confirmed. Biosurfactant showed a reduction in the surface tension to 41 mN/m with a yield concentration of 11.2 g/L. Secondly, its effectiveness was evaluated for the removal of IBU and TCS from the domestic wastewater collected during the dry and rainy seasons. Complete removal of IBU was achieved at 36 h & 6 h and TCS at 6 h & 1 h by crude biosurfactant and biosurfactant at CMC formation for the dry season sample. IBU removal was achieved in 2 h by both crude and biosurfactant at CMC and no TCS was detected in the rainy season sample. Thirdly, biotransformation intermediates of IBU and TCS formed during the application of the biosurfactant and degradation pathways are proposed based on the Liquid Chromatography-Mass Spectrometry (LC-MS) and it indicates that there is no formation of toxic by-products. Based on the results, it is evident that biosurfactant at CMC has performed better for the removal of IBU and TCS than crude biosurfactants without any formation of toxic intermediates. Hence, this study proved to be an eco-friendly, cost-effective and sustainable treatment option for domestic wastewater treatment.

Multifunctional, stable and low-cost lipopeptide biosurfactant produced by Enterobacter cloacae UCP 1597

Caatinga of Pernambuco is an area with a potential richness of microorganisms that produce biosurfactants, which are considered good candidates to replace synthetic surfactants in industrial applications due to their functional stability and low toxicity. In this context, current study aimed to investigate the biosurfactant production by the bacterium Enterobacter cloacae UCP 1597, isolated from the Caatinga soil. First fermentation was carried out in Erlenmeyer flasks containing 100 ml of medium, according to a 2³ full-factorial design (FFD). The results showed higher reduction in surface tension (28.3 mN/m) in condition 2 of the FFD, where dispersion of 38.46 cm2 of burnt motor oil was also verified. Then, a second fermentation was performed in Fernbach flasks, containing 2 L of selected medium, confirming reduction in surface and interfacial tension to 30.5 and 2.3 mN/m, respectively, as well as excellent emulsifying properties. The critical micellar dilution (CMD) of the crude biosurfactant was determined (70%) and its use in phytotoxicity assay verified the absence of toxicity for cabbage seeds. The biomolecule showed a high yield (13.69 g/L) after extraction with ethyl acetate and anionic and lipopeptide nature. The stability in acid pH, high temperature and salinity, showed an acid-resistant, thermostable, and halotolerant biocompound. Thus, this lipopeptide was shown to be a multifunctional biosurfactant, since it not only has excellent surface-active properties, but it is also a good emulsifier, dispersant, and potent agent to germination of cabbage seeds. Hence, is suggested its promising application in industrial activities or environmental processes under adverse conditions.

Assessment of bacterial biosurfactant production and application in Enhanced Oil Recovery (EOR)—A green approach

The present work attempts to investigate production of biosurfactant, in a modified synthetic media, by less explored bacterial strain isolated from iron factory waste effluent from West Bengal, India and its application in enhanced oil recovery operations. The screened isolate was identified to be Pseudomonas mendocina by 16s rDNA gene homology (NCBI-Genebank accession no. MH091331). It showed a high emulsification of 68.3 ± 2.86%, lowering surface tension of water to 31 mN/ m and CMC of 125 mg l −1 and found to be alkalophillic (pH 6 to 12) and halophilic (up to 6% NaCl) in nature. Batch study in optimized media led to maximum yield of 3.09 ± 0.12 g l −1 . Production kinetic analysis by Luedeking Piret model successfully predicted mixed growth associated biosurfactant production with α and β values of 0.73 and 1.60 respectively and a yield coefficient of 0.82. Cost estimation for production of biosurfactant was calculated under lab scale conditions. Characterization of the extracted biosurfactant by anthrone test and Lowry’s protein estimation, TLC, FTIR, H 1 NMR and FESEM revealed the presence of sugar, peptides and lipid moieties. The biosurfactant obtained is extremely robust with high stability, sustaining wide range of pH (4 to 12), salinity (0.5 to 10% NaCl) and temperature (up to 100 °C). Toxicity assessment by Vigna radiata seed bioassay and MTT assay using and HEK293 cell line showed the non-toxic nature of the biosurfactant. Crude biosurfactant could play effective role in oil recovery with 44.3 ± 0.5 % enhanced oil recovery using sand packed column. • Biosurfactant (BS) from newly isolated halo-alkalophillic bacterial strain. • BS production in economically suitable optimized media. • Notably stable BS, tolerating extreme physiological conditions. • BS exhibiting presence of both glycolipids and lipopeptides: blend nature. • Potential use of crude biosurfactant for enhanced oil recovery.

Polycyclic aromatic hydrocarbons remobilization from contaminated porous media by (bio)surfactants washing

Biosurfactants, surface-active agents produced by microorganisms, are increasingly studied for their potential use in soil remediation processes because they are more environmentally friendly than their chemically produced homologues. In this work, we report on the use of a crude biosurfactant produced by a bacterial consortium isolated from a PAHs-contaminated soil, compared with other (bio)surfactants (Tween80, Sodium dodecyl sulfate - SDS, rhamnolipids mix), to wash PAHs from a contaminated porous media. Assays were done using columns filled with sand or sand-clay mixtures (95:5) spiked with four model PAHs. The crude biosurfactant showed less adsorption to the [sand] and the [sand + clay] columns compared to Tween 80, SDS and the rhamnolipid mix. The biosurfactant showed the second best capacity to remove PAHs from the columns (as dissolved and particulate phases), both from [sand] and [sand + clay], after SDS when applied at lower concentrations than the other sufactants. The effluent concentrations of phenanthrene (PHE), pyrene (PYR) and benzo[a]pyrene (BAP) increased in the presence of the crude biosurfactant. Compared to the control experiment using only water, the global PAHs washed mass (amount of PAHs removed from the columns) increased between 9 and 1000 times for PHE and BAP in the [sand] column, and between 55 and 6000 times respectively for PHE and BAP in the [sand + clay] columns. Moreover, in the [sand + clay] columns, leaching of a part of the clays was observed in the SDS and the biosurfactant injections assays. This clay leaching resulted in higher PAHs removal, due not to desorption but rather to particulate transport. In the context of washing PAH-contaminated soils in biopiles or subsurface remediation, our results could help in sizing the remediation approach using an environmental friendly biosurfactant, before a pump-and-treat process.