Production, characterization, evaluation and toxicity assessment of a Bacillus cereus UCP 1615 biosurfactant for marine oil spills bioremediation.

In this study, Bacillus cereus was cultivated in a mineral medium composed of 2% frying oil and 0.12% peptone to produce a biosurfactant. The production was scaled up from flasks to 1.2-, 3.0- and 50-L bioreactors, where surface tension achieved 28.7, 27.5 and 32 mN/m and biosurfactant concentration 4.3, 4.6 and 4.7 g/L, respectively. The biosurfactant was characterized as anionic, while nuclear magnetic resonance, thin-layer chromatography and gas chromatography analyses revealed its lipopeptide nature. Toxicity tests showed survival rates of the fish Poecilia vivipara and the bivalve Anomalocardia brasiliana higher than 90% and 55%, respectively, thus suggesting the use of this biosurfactant in marine environment depollution. Moreover, the biosurfactant stimulated the growth of autochthonous microorganisms independently of the presence of motor oil in bioassays performed in seawater. These results demonstrate that the biosurfactant is biocompatible and has potential for industrial-scale production and application to bioremediation of oil spills-polluted marine environment.

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.

Application of biosurfactants and chitosan in toothpaste formulation.

The aim of the present study was to formulate toothpastes containing biosurfactants and either fungal chitosan or sodium fluoride and evaluate the cytotoxicity, antimicrobial action and inhibition potential against biofilm formed by Streptococcus mutans. Chitosan was extracted from the biomass of the fungus Mucorales. We tested biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus metylotrophicus UCP 1616 (BB) and Candida bombicola URM 3718 (CB). Fractional inhibitory concentration analysis was performed to determine the type of interaction between the compounds. Six toothpaste were prepared, the active ingredients of which were the biosurfactants, chitosan or sodium fluoride. The cytotoxicity tests were performed using the 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. The toothpastes were tested with regard to pH, consistency and foaming capacity. The inhibition of biofilm was investigated by applying the toothpaste to biofilm formed in modified artificial saliva for 24 h at 37 °C in anaerobiosis. All substances had a minimum inhibitory concentration (MIC) for S. mutans. The combinations of CB and PB with chitosan had an additive effect against S. mutans, whereas BB combined with chitosan had an indifferent effect. The toothpastes were non-toxic. The formulations had pH around 9, spreading capacity between 8 and 17 mm and foaming capacity between 63 and 95%. All formulations inhibited the cellular viability of S. mutans in the biofilm, with similar results compared to the commercial toothpaste tested. The present results show that the formulations suggested are promising when compared to a commercial tooth paste.

Mouthwash containing a biosurfactant and chitosan: An eco-sustainable option for the control of cariogenic microorganisms.

The aim of the present study was to determine the antimicrobial action and toxicity of mouthwashes formulated with a biosurfactant, chitosan of a microbial origin and peppermint (Mentha piperita) essential oil (POE). Chitosan was extracted from the biomass of a fungus from the order Mucorales grown in yam bean broth. Three biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus cereus UCP 1615 (BB) and Candida bombicola URM 3718 (CB) were tested. Six mouthwashes were prepared, the active ingredients of which were the biosurfactant, chitosan and POE. The minimum inhibitory concentration (MIC) was determined for the test substances separately, in combinations and in the mouthwash formulas. The toxicity of the mouthwashes was tested using MTT (3-(4,5-dimethylthiazole-2-il)-2,5-diphenyltetrazolium bromide) for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. All substances tested had a MIC for cariogenic microorganisms. The combinations of the CB and PB biosurfactants with chitosan demonstrated an additive effect on the majority of microorganisms tested. The toxicity of the mouthwashes was significantly lower than that of the commercial mouthwash. The present findings demonstrate that mouthwashes containing natural products constitute a safe, effective, natural alternative to commercially available mouthwashes for the control of oral microorganisms.

Application of a low-cost biosurfactant in heavy metal remediation processes.

The industrial interest in microbial surfactants has intensified in recent years due to the characteristics of these compounds, such as biodegradability, low toxicity, and effectiveness in removing heavy metals and hydrophobic organic compounds from soil and water. This paper describes the production of a biosurfactant by the yeast Candida tropicalis grown in distilled water with 2.5% molasses, 2.5% frying oil and 4% corn steep liquor. The production of the biosurfactant reached 27 g/l in a 50-l bioreactor with a surface tension of 30 mN/m. Surface tension and engine oil emulsification assays demonstrated the stability of biosurfactant under extreme conditions of temperature and pH as well as in the presence of NaCl. Chemical structures of the biosurfactant were identified using GC-MS and NMR. The isolated biosurfactant was characterised as an anionic molecule capable of reducing the surface tension of water from 70 to 30 mN/m at 0.5% of the critical micelle concentration, with no toxic effects on plant seeds or brine shrimp. In tests involving both the crude and isolated biosurfactant for the removal of heavy metals from contaminated sand under dynamic conditions, the removal rates for Zn and Cu ranged from 30 to 80%, while the best removal rate for Pb was 15%. Tests in packed columns also confirmed the ability of biosurfactant to remove Cu and Zn at rates ranging from 45 to 65%. However, lead was not removed under static conditions. The removal kinetics demonstrated that 30 min was sufficient for the removal of metals and a single washing with the biosurfactant achieved greater removal efficiency. The use of the biosurfactant led to a significant reduction in the electrical conductivity of solutions containing heavy metals. The present findings as well as a brief economic analysis suggest the great potential of this agent for industrial remediation processes of soil and water polluted with inorganic contaminants.

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.

Use of bacterial biosurfactants as natural collectors in the dissolved air flotation process for the treatment of oily industrial effluent.

The aim of the present study was to investigate the separation of oil from water using a bench-scale DAF prototype with the addition of biosurfactants isolated from Pseudomonas cepacia CCT6659 and Bacillus cereus UCP1615. The best operating conditions for the DAF prototype were determined using a central composite rotatable design. The results demonstrated that the biosurfactants from P. cepacia and B. cereus increased the oil separation efficiency from 53.74% (using only microbubbles) to 94.11 and 80.01%, respectively. The prediction models for both DAF-biosurfactant systems were validated, showing an increase in the efficiency of the DAF process from 53.74% to 98.55 and 70.87% using the formulated biosurfactants from P. cepacia and B. cereus, respectively. The biosurfactant from P. cepacia was selected as the more promising product and used for the treatment of oily effluent from a thermoelectric plant, achieving removal rates ranging between 75.74 (isolated biosurfactant) and 95.70% (formulated biosurfactant), respectively.

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.

Yeasts and bacterial biosurfactants as demulsifiers for petroleum derivative in seawater emulsions.

Oil sludge or waste generated in transport, storage or refining forms highly stable mixtures due to the presence and additives with surfactant properties and water forming complex emulsions. Thus, demulsification is necessary to separate this residual oil from the aqueous phase for oil processing and water treatment/disposal. Most used chemical demulsifiers, although effective, are environmental contaminants and do not meet the desired levels of biodegradation. We investigated the application of microbial biosurfactants as potential natural demulsifiers of petroleum derivatives in water emulsions. Biosurfactants crude extracts, produced by yeasts (Candida guilliermondii, Candida lipolytica and Candida sphaerica) and bacteria (Pseudomonas aeruginosa, Pseudomonas cepacia and Bacillus sp.) grown in industrial residues, were tested for demulsification capacity in their crude and pure forms. The best results obtained were for bacterial biosurfactants, which were able to recover about 65% of the seawater emulsified with motor oil compared to 35-40% only for yeasts products. Biosurfactants were also tested with oil-in-water (O/W) and water-in-oil (W/O) kerosene model emulsions. No relationship between interfacial tension, cell hydrophobicity and demulsification ratios was observed with all the biosurfactants tested. Microscopic illustrations of the emulsions in the presence of the biosurfactants showed the aspects of the emulsion and demulsification process. The results obtained demonstrate the potential of these agents as demulsifiers in marine environments.

Candida lipolytica UCP0988 Biosurfactant: Potential as a Bioremediation Agent and in Formulating a Commercial Related Product.

The aim of the present study was to investigate the potential application of the biosurfactant from Candida lipolytica grown in low-cost substrates, which has previously been produced and characterized under optimized conditions as an adjunct material to enhance the remediation processes of hydrophobic pollutants and heavy metals generated by the oil industry and propose the formulation of a safe and stable remediation agent. In tests carried out with seawater, the crude biosurfactant demonstrated 80% oil spreading efficiency. The dispersion rate was 50% for the biosurfactant at a concentration twice that of the CMC. The biosurfactant removed 70% of motor oil from contaminated cotton cloth in detergency tests. The crude biosurfactant also removed 30-40% of Cu and Pb from standard sand, while the isolated biosurfactant removed ~30% of the heavy metals. The conductivity of solutions containing Cd and Pb was sharply reduced after biosurfactants' addition. A product was prepared through adding 0.2% potassium sorbate as preservative and tested over 120 days. The formulated biosurfactant was analyzed for emulsification and surface tension under different pH values, temperatures, and salt concentrations and tested for toxicity against the fish Poecilia vivipara. The results showed that the formulation had no toxicity and did not cause significant changes in the tensoactive capacity of the biomolecule while maintaining activity demonstrating suitability for potential future commercial product formulation.

Response Surface Methodology for Optimizing the Production of Biosurfactant by Candida tropicalis on Industrial Waste Substrates.

Biosurfactant production optimization by Candida tropicalis UCP0996 was studied combining central composite rotational design (CCRD) and response surface methodology (RSM). The factors selected for optimization of the culture conditions were sugarcane molasses, corn steep liquor, waste frying oil concentrations and inoculum size. The response variables were surface tension and biosurfactant yield. All factors studied were important within the ranges investigated. The two empirical forecast models developed through RSM were found to be adequate for describing biosurfactant production with regard to surface tension (R2 = 0.99833) and biosurfactant yield (R2 = 0.98927) and a very strong, negative, linear correlation was found between the two response variables studied (r = -0.95). The maximum reduction in surface tension and the highest biosurfactant yield were 29.98 mNm-1 and 4.19 gL-1, respectively, which were simultaneously obtained under the optimum conditions of 2.5% waste frying oil, 2.5%, corn steep liquor, 2.5% molasses, and 2% inoculum size. To validate the efficiency of the statistically optimized variables, biosurfactant production was also carried out in 2 and 50 L bioreactors, with yields of 5.87 and 7.36 gL-1, respectively. Finally, the biosurfactant was applied in motor oil dispersion, reaching up to 75% dispersion. Results demonstrated that the CCRD was suitable for identifying the optimum production conditions and that the new biosurfactant is a promising dispersant for application in the oil industry.

Production of Biosurfactants by Pseudomonas Species for Application in the Petroleum Industry.

The production of surfactants by microorganisms has become an attractive option in the treatment of oil-contaminated environments because biosurfactants are biodegradable and less toxic than synthetic surfactants, although production costs remain high. With the aim of reducing the cost of biosurfactant production, three strains of Pseudomonas (designated P1, P2, and P3) were cultivated in a low-cost medium containing molasses and corn steep liquor as substrates. Following the selection of the best producer (P3), a rotational central composite design (RCCD) was used to determine the influence of substrates concentration on surface tension and biosurfactant yield. The biosurfactant reduced the surface tension of water to 27.5 mN/m, and its CMC was determined to be 600 mg/L. The yield was 4.0 g/L. The biosurfactant demonstrated applicability under specific environmental conditions and was able to remove 80 to 90% of motor oil adsorbed to sand. The properties of the biosurfactant suggest its potential application in bioremediation of hydrophobic pollutants.

Biosurfactants: Promising Molecules for Petroleum Biotechnology Advances.

The growing global demand for sustainable technologies that improves the efficiency of petrochemical processes in the oil industry has driven advances in petroleum biotechnology in recent years. Petroleum industry uses substantial amounts of petrochemical-based synthetic surfactants in its activities as mobilizing agents to increase the availability or recovery of hydrocarbons as well as many other applications related to extraction, treatment, cleaning, and transportation. However, biosurfactants have several potential applications for use across the oil processing chain and in the formulations of petrochemical products such as emulsifying/demulsifying agents, anticorrosive, biocides for sulfate-reducing bacteria, fuel formulation, extraction of bitumen from tar sands, and many other innovative applications. Due to their versatility and proven efficiency, biosurfactants are often presented as valuable versatile tools that can transform and modernize petroleum biotechnology in an attempt to provide a true picture of state of the art and directions or use in the oil industry. We believe that biosurfactants are going to have a significant role in many future applications in the oil industries and in this review therefore, we highlight recent important relevant applications, patents disclosures and potential future applications for biosurfactants in petroleum and related industries.

Formulation of a Commercial Biosurfactant for Application as a Dispersant of Petroleum and By-Products Spilled in Oceans.

Oil spills in oceans cause irreparable damage to marine life and harm the coastal populations of affected areas. It is therefore fundamental to develop treatment strategies for such spills. Currently, chemical dispersants have been used during oil spills, although these agents have been increasingly restricted due to their toxic potential. Thus, the aim of the present study was to formulate a biodegradable commercial biosurfactant for application as a dispersant. Biosurfactants are scientifically known biomolecules produced by microorganisms capable of allowing water-oil interaction. Thus, a biosurfactant was produced by the yeast Candida bombicola URM 3718 cultivated in industrial waste and formulated with the addition of a potassium sorbate preservative for fractionated sterilization (tyndallization) and the combination of fluent vaporization with the preservative. After formulation, samples were stored for 120 days, followed by surface tension, emulsification and oil dispersant tests in sea water. The results were promising for the biosurfactant formulated with the preservative, which demonstrated stability and an absence of toxicity in experiments with a marine indicator. The commercial biosurfactant was tested at different pH values, temperatures and in the presence of salt, demonstrating potential industrial application at a cost compatible with the environmental field. The formulation process developed in this research was patented in the Brazilian National Intellectual Property Institute (patent number BR1020140179631).

Biosurfactants: Multifunctional Biomolecules of the 21st Century.

In the era of global industrialisation, the exploration of natural resources has served as a source of experimentation for science and advanced technologies, giving rise to the manufacturing of products with high aggregate value in the world market, such as biosurfactants. Biosurfactants are amphiphilic microbial molecules with hydrophilic and hydrophobic moieties that partition at liquid/liquid, liquid/gas or liquid/solid interfaces. Such characteristics allow these biomolecules to play a key role in emulsification, foam formation, detergency and dispersal, which are desirable qualities in different industries. Biosurfactant production is considered one of the key technologies for development in the 21st century. Besides exerting a strong positive impact on the main global problems, biosurfactant production has considerable importance to the implantation of sustainable industrial processes, such as the use of renewable resources and "green" products. Biodegradability and low toxicity have led to the intensification of scientific studies on a wide range of industrial applications for biosurfactants in the field of bioremediation as well as the petroleum, food processing, health, chemical, agricultural and cosmetic industries. In this paper, we offer an extensive review regarding knowledge accumulated over the years and advances achieved in the incorporation of biomolecules in different industries.

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.

Applications of biosurfactants in the petroleum industry and the remediation of oil spills.

Petroleum hydrocarbons are important energy resources. However, petroleum is also a major pollutant of the environment. Contamination by oil and oil products has caused serious harm, and increasing attention has been paid to the development and implementation of innovative technologies for the removal of these contaminants. Biosurfactants have been extensively used in the remediation of water and soil, as well as in the main stages of the oil production chain, such as extraction, transportation, and storage. This diversity of applications is mainly due to advantages such as biodegradability, low toxicity and better functionality under extreme conditions in comparison to synthetic counterparts. Moreover, biosurfactants can be obtained with the use of agro-industrial waste as substrate, which helps reduce overall production costs. The present review describes the potential applications of biosurfactants in the oil industry and the remediation of environmental pollution caused by oil spills.

Characterization of a biosurfactant produced by Pseudomonas cepacia CCT6659 in the presence of industrial wastes and its application in the biodegradation of hydrophobic compounds in soil.

The bacterium Pseudomonas cepacia CCT6659 cultivated with 2% soybean waste frying oil and 2% corn steep liquor as substrates produced a biosurfactant with potential application in the bioremediation of soils. The biosurfactant was classified as an anionic biomolecule composed of 75% lipids and 25% carbohydrates. Characterization by proton nuclear magnetic resonance ((1)H and (13)C NMR) revealed the presence of carbonyl, olefinic and aliphatic groups, with typical spectra of lipids. Four sets of biodegradation experiments were carried out with soil contaminated by hydrophobic organic compounds amended with molasses in the presence of an indigenous consortium, as follows: Set 1-soil+bacterial cells; Set 2-soil+biosurfactant; Set 3-soil+bacterial cells+biosurfactant; and Set 4-soil without bacterial cells or biosurfactant (control). Significant oil biodegradation activity (83%) occurred in the first 10 days of the experiments when the biosurfactant and bacterial cells were used together (Set 3), while maximum degradation of the organic compounds (above 95%) was found in Sets 1-3 between 35 and 60 days. It is evident from the results that the biosurfactant alone and its producer species are both capable of promoting biodegradation to a large extent.