Microbial Biosurfactant: Candida bombicola as a Potential Remediator of Environments Contaminated by Heavy Metals.

Industrial interest in surfactants of microbial origin has intensified recently due to the characteristics of these compounds, such as biodegradability and reduced toxicity, and their efficiency in removing heavy metals and hydrophobic organic compounds from soils and waters. The aim of this study was to produce a biosurfactant using Candida bombicola URM 3712 in a low-cost medium containing 5.0% molasses, 3.0% corn steep liquor and 2.5% residual frying oil for 144 h at 200 rmp. Measurements of engine oil tension and emulsification were made under extreme conditions of temperature (0 °C, 5 °C, 70 °C, 100 °C and 120 °C), pH (2-12) and NaCl concentrations (2-12), demonstrating the stability of the biosurfactant. The isolated biosurfactant was characterized as an anionic molecule with the ability to reduce the surface tension of water from 72 to 29 mN/m, with a critical micellar concentration of 0.5%. The biosurfactant had no toxic effect on vegetable seeds or on Eisenia fetida as a bioindicator. Applications in the removal of heavy metals from contaminated soils under dynamic conditions demonstrated the potential of the crude and isolated biosurfactant in the removal of Fe, Zn and Pb with percentages between 70 and 88%, with the highest removal of Pb being 48%. The highest percentage of removal was obtained using the cell-free metabolic liquid, which was able to remove 48, 71 and 88% of lead, zinc and iron from the soil, respectively. Tests in packed columns also confirmed the biosurfactant's ability to remove Fe, Zn and Pb between 40 and 65%. The removal kinetics demonstrated an increasing percentage, reaching removal of 50, 70 and 85% for Pb, Zn and Fe, respectively, reaching a greater removal efficiency at the end of 24 h. The biosurfactant was also able to significantly reduce the electrical conductivity of solutions containing heavy metals. The biosurfactant produced by Candida bombicola has potential as an adjuvant in industrial processes for remediating soils and effluents polluted by inorganic contaminants.

Treatment of oily effluent using a low-cost biosurfactant in a flotation system.

Fuel and lubricating oil leaks produce an oily wastewater that creates an environmental problem for industries. Dissolved air flotation (DAF) has been successfully employed for the separation of oily contaminants. Collectors constitute an auxiliary tool in the DAF process that enhances the separation efficiency by facilitating the adhesion of the contaminant particles. The use of biosurfactants as collectors is a promising technology in flotation processes, as these biomolecules are biodegradable and non-toxic. In the present study, a biosurfactant was produced from the bacteria Pseudomonas aeruginosa UCP 0992 cultivated in 0.5% corn steep liquor and 4.0% vegetable oil residue in a bioreactor at 225 rpm for 120 h, resulting in a surface tension of 26.5 mN/m and a yield of 26 g/L. The biosurfactant demonstrated stability when exposed to different temperatures, heating times, pH values and salt and was characterised as a glycolipid with a critical micelle concentration of 600 mg/L. A central composite rotatable design was used to evaluate the effect of the crude biosurfactant added to a laboratory DAF prototype on the removal efficiency of motor oil. The isolated and formulated forms of the biosurfactant were also tested in the prototype after the optimisation of the operational conditions. The results demonstrated that all forms of the biosurfactant increased the oil separation efficiency of the DAF process by 65 to 95%. In conclusion, the use of biosurfactants is a promising alternative as an auxiliary tool in flotation processes for the treatment of oily waters generated by industrial activities.

Production and formulation of a new low-cost biosurfactant to remediate oil-contaminated seawater.

The aim of the present study was to produce biosurfactants using three bacterial strains (Pseudomonas cepacia CCT6659, Bacillus methylotrophicus UCP 1616 and Bacillus cereus UCP 1615) cultivated in mineral medium containing different carbon (glucose, sucrose, molasses and waste frying oil) and nitrogen [NH4NO3, (NH4)2SO4, peptone, yeast extract and corn steep liquor] sources. B. cereus stood out as the best biosurfactant producer when inoculated with a 1.5% cell suspension and cultivated at 28 °C and 200 rpm in 2.0% molasses and 1.0% corn steep liquor for 48 h. Under these conditions, medium surface tension was reduced to 26.2 ± 0.2 mN/m, and biosurfactant concentration achieved 2.05 ± 0.32 g/L. The biosurfactant showed a critical micelle concentration of 0.90 ± 0.05 g/L, proved to be highly stable in wide ranges of pH, salt concentration and heating temperature, and exerted low toxicity to larvae of Artemia salina as a marine environmental bioindicator. Structural characterisation of biosurfactant suggested a lipopeptide composition. The biotensioactive agent was shown to effectively remove motor oil adsorbed to marine rock (91.0 ± 0.4%) and to disperse it in seawater (70.0 ± 0.4%). The biosurfactant formulated with 0.2% potassium sorbate demonstrated considerable potential for application in the petroleum industry, where it could be successfully used as a commercial product to mobilize oil in marine environments.

Production of a biosurfactant from Bacillus methylotrophicus UCP1616 for use in the bioremediation of oil-contaminated environments.

The aim of the present study was to produce a microbial biosurfactant for use in the bioremediation of environments contaminated with petroleum products. Bacillus methylotrophicus was isolated from seawater taken from a port area and cultivated using industrial waste as substrate (corn steep liquor and sugarcane molasses [both at 3%]). Surface tension measurements and motor oil emulsification capacity were used for the evaluation of the production of the biosurfactant, which demonstrated stability in a broad range of pH and temperature as well as a high concentration of saline, with the reduction of the surface tension of water to 29 mN/m. The maximum concentration of biosurfactant (10.0 g/l) was reached after 144 h of cultivation. The biosurfactant was considered to be a lipopeptide based on the results of proton nuclear magnetic resonance and Fourier transformed infrared spectroscopy. The tests demonstrated that the biosurfactant is innocuous and has potential for the bioremediation of soil and water contaminated by petroleum products. Thus, the biosurfactant described herein has a low production cost and can be used in environmental processes.

Recovery of contaminated marine environments by biosurfactant-enhanced bioremediation.

The need to remediate areas contaminated by petroleum products has led to the development of novel technologies for treating such contaminants in a non-conventional manner, that is, without the use of chemical or physical methods. Biosurfactants are amphipathic biomolecules produced by microorganisms that can be used in bioremediation processes in environments contaminated by petroleum products due to their excellent tensioactive properties. The aim of the present study was to produce a biosurfactant from Pseudomonas aeruginosa UCP 0992 cultivated in 0.5% corn steep liquor and 4.0% vegetable oil residue in a 1.2-L bioreactor employing a central composite rotatable design to optimize the cultivation conditions for maximum yield. The best results were achieved with aeration rate of 1.0 vvm and 3.0% inoculum at 225 rpm for 120 h, resulting in a surface tension of 26.5 mN/m and a biosurfactant yield of 26 g/L. Kinetic and static assays were then performed with the biosurfactant for the removal of motor oil adsorbed to sand, with removal rates around 90% and 80%, respectively, after 24 h. Oil degradation experiments with the bacterium and the combination of the bacterium and biosurfactant were also conducted to simulate the bioremediation process in sand and seawater samples (duration: 75 and 30 days, respectively). In both cases, oil degradation rates were higher than 90% in the presence of the biosurfactant and the producing species, indicating the potential of the biomolecule as an adjuvant in petroleum decontamination processes in the marine environment.

Saponins and microbial biosurfactants: Potential raw materials for the formulation of cosmetics.

The cosmetic industry is currently one of the fasting growing sections of the economy in many countries. The recent tendency toward the use of cosmetics of a natural origin has driven the industry to seek alternatives to synthetic components in the formulation of products. Biosurfactants are natural compounds that have considerable potential for application in the formulation of safe, effective cosmetics as a replacement for commonly used chemical tensioactive agents. The present review provides essential information on the physicochemical and biological properties of saponins and microbial biosurfactants employed in cosmetic products, with a focus on the use of these natural compounds in shampoos, addressing the current state of research and patents involving biosurfactants for this purpose. The challenges and prospects of this cosmetic application are also discussed. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1482-1493, 2018.

Application of bacterial and yeast biosurfactants for enhanced removal and biodegradation of motor oil from contaminated sand

Background This study investigated the potential application of two biosurfactants for enhanced removal capability and biodegradation of motor oil contaminated sand under laboratory conditions. The biosurfactants were produced by the yeast Candida sphaerica and by the bacterium Bacillus sp. cultivated in low-cost substrates. The ability of removing motor oil from soil by the two biosurfactants was identified and compared with that of the synthetic surfactants Tween 80 and Triton X-100. Results Both crude and isolated biosurfactants showed excellent effectiveness on motor oil removal from contaminated sand under kinetic conditions (70-90%), while the synthetic surfactants removed between 55 and 80% of the oil. A contact time of 5-10 min under agitation seemed to be enough for oil removal with the biosurfactants and synthetic surfactants tested. The crude and the isolated biosurfactant from C. sphaerica were able to remove high percentages of motor oil from packed columns (around 90%) when compared to the biosurfactant from Bacillus sp. (40%). For the degradation experiments conducted in motor oil contaminated sand enriched with sugar cane molasses, however, oil degradation reached almost 100% after 90 d in the presence of Bacillus sp. cells, while the percentage of oil degradation did not exceed 50% in the presence of C. sphaerica. The presence of the biosurfactants increased the degradation rate in 10-20%, especially during the first 45 d, indicating that biosurfactants acted as efficient enhancers for hydrocarbon biodegradation. Conclusions The results indicated the biosurfactants enhancing capability on both removal and rate of motor oil biodegradation in soil systems.

Formulation of mayonnaise with the addition of a bioemulsifier isolated from Candida utilis.

Biosurfactants have a number of industrial applications due their diverse properties, such as emulsification, foaming, wetting, and surface activity. The aim of the present study was to produce a biosurfactant from Candida utilis and employ it in the formulation of a mayonnaise. The biosurfactant was produced in a mineral medium supplemented with glucose and canola waste frying oil at 150 rpm for 88 h. The product was biologically tested on rats and in different formulations of mayonnaise, which were submitted to microbiological evaluations. The biosurfactant was added to the diet of the rats for 21 days. Greater consumption was found of the experimental diet. Moreover, no changes were found in the liver or kidneys of the animals, demonstrating the absence of a toxic effect from the biosurfactant. Six different formulations of mayonnaise were prepared and tested regarding stability with the addition of carboxymethyl cellulose and guar gum (combined and isolated) after 30 days of refrigeration. The most stable formulation with the best quality was obtained with combination of guar gum and the isolated biosurfactant, with an absence of pathogenic microorganisms. In conclusion, the potential and innocuousness of the biosurfactant isolated from C. utilis indicates its safe use in food emulsions.

Production of a bioemulsifier with potential application in the food industry.

Biosurfactants are of considerable interest due to their biodegradability, low degree of toxicity, and diverse applications. However, the high production costs involved in the acquisition of biosurfactants underscore the need for optimization of the production process to enable viable application on an industrial scale. The aims of the present study were to select a species of Candida that produces a biosurfactant with the greatest emulsifying potential and to investigate the influence of components of the production medium and cultivation conditions. Candida utilis achieved the lowest surface tension (35.53 mN/m), best emulsification index (73%), and highest yield (12.52 g/l) in a medium containing waste canola frying oil as the carbon source and ammonium nitrate as the nitrogen source. The best combination of medium components and cultivation conditions was 6% (w/v) glucose, 6% (w/v) waste canola frying oil, 0.2% (w/v) ammonium nitrate, 0.3% (w/v) yeast extract, 150 rpm, 1% inoculum (w/v), and 88 h of fermentation. The greatest biosurfactant production and the lowest surface tension were achieved in the first 24 h of production, and the maximum biomass production was recorded at 72 h. The biosurfactant produced from C. utilis under the conditions investigated in the present study has a potential to be a bioemulsifier for application in the food industry.

Characterisation, surface properties and biological activity of a biosurfactant produced from industrial waste by Candida sphaerica UCP0995 for application in the petroleum industry.

The development of less toxic, biodegradable, surfactants, such as biosurfactants, is a key strategy for acquiring environmentally friendly compounds. The aim of the present study was to employ an optimised medium containing 9% ground nut oil refinery residue and 9% corn steep liquor for the production of a biosurfactant by Candida sphaerica. Fermentation was carried out at 28 °C and 200 rpm for 144 h. Biosurfactant yield was 9 g/l. The biosurfactant reduced the surface tension of the medium to 25 mN/m, with a critical micelle concentration of 0.025%. The product demonstrated stability with regard to surface tension reduction and emulsification in a range of temperatures (5-120 °C) and pH values (2-12) as well as tolerance to high concentrations of NaCl (2-10%). Hydrophobicity tests indicate two possible insoluble substrate uptake mechanisms: direct interfacial uptake and biosurfactant-mediated transfer (cell contact with emulsified or solubilised hydrocarbons). The biosurfactant was characterised as an anionic glycolipid consisting of 70% lipids and 15% carbohydrates and demonstrated no toxicity to the microcrustacean Artemia salina or the vegetables Brassica oleracea, Solanum gilo, Lactuca sativa L. and Brassica oleracea L. The biosurfactant recovered 95% of motor oil adsorbed to a sand sample, demonstrating considerable potential for use in bioremediation processes, especially in the petroleum industry.

Economic optimized medium for tensio-active agent production by Candida sphaerica UCP0995 and application in the removal of hydrophobic contaminant from sand.

Statistical experimental designs and response surface methodology were employed to optimize the concentrations of agroindustrial residues as soybean oil (SORR) from refinery, and corn steep liquor (CSL) from corn industry, for tensio-active agent produced by Candida sphaerica UCP 0995. Three 2(2) full factorial design were applied sequentially to investigate the effects of the concentrations and interactions of soybean oil refinery residue and corn steep liquor on the surface tension of free-cell culture broth for 144 h. Two 2(2) central composite designs and response surface methodology were adopted to derive a statistical model to measure the effect of SORR and CSL on the surface tension of the free-cell culture broth for 144 h. The regression equation obtained from the experimental data using a central composite design was solved, and by analyzing the response surface contour plots, the optimal concentrations of the constituents of the medium were determined: 8.63% v/v (≅9% v/v) of SORR and 8.80% v/v (≅9% v/v) CSL. The minimum surface tension predicted and experimentally confirmed was 25.25 mN/m. The new biosurfactant, denominated Lunasan, recovered 95% of motor oil adsorbed in a sand sample, thus showing great potential for use in bioremediation processes, especially in the petroleum industry.

Evaluation antimicrobial and antiadhesive properties of the biosurfactant Lunasan produced by Candida sphaerica UCP 0995.

Different groups of biosurfactants exhibit diverse properties and display a variety of physiological functions in producer microorganisms; these include enhancing the solubility of hydrophobic/water-insoluble compound, heave metal binding, bacterial pathogenesis, cell adhesion and aggregation, quorum sensing and biofilm formation. Candida sphaerica was grown in a low cost medium, consisting of distilled water supplemented with 9% refinery residue of soybean oil and 9% corn steep liquor, for 144 h at 28°C and 150 rpm. The cell-free supernatant obtained at the end of the experiments was submitted to extraction, and afterward the biosurfactant was isolated using methanol with a yield of 9 g l(-1). The critical micelle concentration of the biosurfactant was found to be 0.25 mg ml(-1) with a surface tension of 25 mN m(-1). Several concentrations of the biosurfactant (0.625-10 mg ml(-1)) were used to evaluate its antimicrobial and antiadhesive activities against a variety of microorganisms. The biosurfactant showed antimicrobial activity against Streptococcus oralis (68%), Candida albicans (57%), and Staphylococcus epidermidis(57.6%) for the highest concentration tested. Furthermore, the biosurfactant at a concentration of 10 mg ml(-1) inhibited the adhesion between 80 and 92% of Pseudomonas aeruginosa, Streptococcus agalactiae, Streptococcus sanguis12. Inhibition of adhesion with percentages near 100% occurred for the higher concentrations of biosurfactant used. Results gathered in this study point to a potential use of the biosurfactant in biomedical applications.

Studies of the cell surface properties of Candida species and relation to the production of biosurfactants for environmental applications.

In practical bioremediation of petroleum pollution, treatment systems often use soil, sand, and other aquifer porous media besides water solutions. The distribution of the microbial cell also plays an important role in the whole process of bioremediation; therefore, the adhesion ability of cells to porous media is one of the key factors influencing the efficiency of treatment. The probable modes of hydrocarbon uptake in cells of Candida were studied based on data for cell hydrophobicity, emulsifying activity, surface tension, and interfacial tension of the cell-free culture medium. Six Candida strains were cultivated in insoluble and soluble substrates for 144 h, including n-hexadecane, soybean oil, ground-nut oil refinery residue, corn steep liquor, and glucose. The results obtained showed the potential of yeasts for application in the removal of hydrophobic compounds. Depending the strain and substrate used the adhesion ability of yeast cells and the production of surfactants and emulsifiers can take place simultaneously, thus increasing the efficiency of bioremediation treatment of petroleum pollution. The application of crude biosurfactants separated from the yeast cells was also demonstrated by tests of removal of petroleum and the derivate motor oil adsorbed in sand samples. Biosurfactants produced in low-cost medium were able to remove 90% of the hydrophobic contaminants.

Experimental design for the production of tensio-active agent by Candida lipolytica.

The strategy of optimization using sequential factorial design was employed to enhance the tensio-active emulsifying agent produced by Candida lipolytica using soybean oil refinery residue as substrate. A full factorial design was used to evaluate the impact of three fermentation factors-amounts of refinery residue, glutamic acid and yeast extract. This allowed exclusion of the yeast extract. Full factorials designs were then sequentially used to optimize the levels of the residue and glutamic acid. The surface tension value was finally reduced to 25.29 mN/m. The maximum emulsifier activity using different substrates was within 40 h of cultivation. The surface tension of the cell-free broth containing the biosurfactant remained very stable during exposure to a wide range of pH (2-12), temperatures (0-120 degrees C) and salinity (2-10% NaCl). The combination of an industrial waste and a cheap substrate therefore seems to be very promising for the low-cost production of potent biosurfactant.

Co-utilization of canola oil and glucose on the production of a surfactant by Candida lipolytica.

Candida lipolytica synthesized a surfactant in a cultivation medium supplemented with canola oil and glucose as carbon sources. Measurements of biosurfactant production and surface tension indicated that the biosurfactant was produced at 48 h of fermentation. The surface-active species is constituted by the protein-lipid-polysaccharide complex in nature. The cell-free broth was particularly influenced by the addition of salt, the pH and temperature depending on the emulsified substrate (hexadecane or a vegetable oil). After comparison between ethyl acetate and mixtures of chloroform and methanol as solvent systems for surfactant recovery, it was found that ethyl acetate was able to extract crude surfactant material with high product recovery (8.0 g/L). The isolated biosurfactant decreased the surface tension to values of 30 mN/m at the critical micelle concentration. Emulsification properties of the biosurfactant produced were compared to those of commercial emulsifiers and other microbial surfactants.