C. Calvo

Application of bioemulsifiers in soil oil bioremediation processes. Future prospects

Biodegradation is one of the primary mechanisms for elimination of petroleum and other hydrocarbon pollutants from the environment. It is considered an environmentally acceptable way of eliminating oils and fuel because the majority of hydrocarbons in crude oils and refined products are biodegradable. Petroleum hydrocarbon compounds bind to soil components and are difficult to remove and degrade. Bioemulsifiers can emulsify hydrocarbons enhancing their water solubility and increasing the displacement of oily substances from soil particles. For these reasons, inclusion of bioemulsifiers in a bioremediation treatment of a hydrocarbon polluted environment could be really advantageous. There is a useful diversity of bioemulsifiers due to the wide variety of producer microorganisms. Also their chemical compositions and functional properties can be strongly influenced by environmental conditions. The effectiveness of the bioemulsifiers as biostimulating agent in oil bioremediation processes has been demonstrated by several authors in different experimental assays. For example, they have shown to be really efficient in combination with other products frequently used in oil bioremediation such as they are inorganic fertilizer (NPK) and oleophilic fertilizer (i.e. S200C). On the other hand, the bioemulsifiers have shown to be more efficient in the treatment of soil with high percentage of clay. Finally, it has been proved their efficacy in other biotechnological processes such as in situ treatment and biopiles. This paper reviews literature concerning the application of bioemulsifiers in the bioremediation of soil polluted with hydrocarbons, and summarizes aspects of the current knowledge about their industrial application in bioremediation processes.

Characterization of exopolysaccharides produced by 19 halophilic strains of the species Halomonas eurihalina

The formation, chemical composition and rheological properties of the exopolysaccharides (EPS) produced by 19 strains belonging to Halomonas eurihalina have been compared in two different culture media. Our aim was to screen several strains isolated from saline soils to select those producing maximum EPS yield and good rheological properties. We found that MY medium was best for the production of EPS in all the strains studied. Maximum EPS production was 1.6 g l 1 with strain H212 grown in this medium. The pattern of the chemical composition of the polysaccharides was affected by the strain in question and by the culture medium. All EPS studied had an unusually high sulphate content. Furthermore, the exopolymer from strain H96 contained significant amounts of uronic acid. EPS from strain H96, cultivated in defined NH medium, behaved in an interesting way rheologically; when the pH of the polymer solution was decreased to 3.0 a gel with a viscosity of 30 000 cP formed.

Exopolysaccharide production byVolcaniella eurihalina

The application of microbial extracellular polymers began in the 1960s, and since then there has been a remarkable increase in their commercial use. They are used in the food, textile, pharmaceutical, agricultural, paint and petroleum industries.Volcaniella eurihalina, a moderately halophilic eubacterium, produces an extracellular polysaccharide whose physical and chemical properties could be of interest for various industrial applications. The aim of this investigation is to analyze the different environmental parameters which influence the production of polysaccharide, and to study its chemical composition and the rheological properties of its solutions.

Effect of cations, pH and sulfate content on the viscosity and emulsifying activity of the Halomonas eurihalina exopolysaccharide

The effects of monovalent and divalent cations on the rheological behavior of Halomonas eurihalina exopolysaccharide (EPS) were studied. Sodium, potassium, magnesium and calcium were added and the relative abilities to increase viscosity were as follows: KCl > NaCl > MgCl2 > CaCl2. The highest viscosity value was measured in acidic 10−4 M KCl, in which a gel formed. A loss of sulfate content seemed to correlate with the increase of viscosity. H. eurihalina produced EPS in all growth media. Addition of hydrophobic substrates to culture media produced changes in chemical composition and emulsifying activity of the EPS. Xylene was the most effectively emulsified substance and the EPS produced on tetradecane and on corn oil the most active emulsifier.

Yield production, chemical composition, and functional properties of emulsifier H28 synthesized by Halomonas eurihalina strain H-28 in media containing various hydrocarbons.

Abstract.Halomonas eurihalina strain H-28 is a moderately halophilic bacterium that produces an extracellular polysaccharide not only in media with glucose but also in media supplemented with hydrocarbons (n-tetradecane, n-hexadecane, n-octane, xylene, mineral light oil, mineral heavy oil, petrol, or crude oil). In this study we investigated yield production, chemical composition, viscosity, and emulsifying activity of exopolysaccharides (EPS) extracted from the different media used. The largest amounts of biopolymer were synthesized in media with glucose and n-hexadecane. Chemical composition varied with culture conditions; thus EPS from cultures grown in the presence of hydrocarbons had lower contents of carbohydrates and proteins than EPS from media with glucose. However, the percentages of uronic acids, acetyls, and sulfates were always higher than glucose EPS. Crude oil was the substrate most effectively emulsified. All EPS were capable of emulsifying crude oil more efficiently than the three control surfactants tested (Tween 20, Tween 80, and Triton X-100). All polymers gave low viscosity solutions. EPS H28 could be attractive for application in the oil industry and/or in bioremediation processes, bearing in mind not only its functional properties, but also the capacity of producer strain H-28 to grow in the presence of high salt concentrations and oil substrates.

Characteristics of bioemulsifiers synthesised in crude oil media by Halomonas eurihalina and their effectiveness in the isolation of bacteria able to grow in the presence of hydrocarbons

Abstract.Halomonas eurihalina strains F2–7, H28, H96, H212 and H214 were capable of producing large amounts of exopolysaccharides (EPS) in MY medium with added crude oil. The biopolymers showed lower carbohydrate and protein content than those synthesised in control medium without oil. Nevertheless, the percentages of uronic acids, acetyls and sulphates were higher. The emulsifying activity of biopolymers was measured; crude oil was the substrate most efficiently emulsified. Furthermore, all the EPS tested emulsified higher volumes of crude oil than the commercial surfactants used as controls. We have also proved the effectiveness of both Halomonas eurihalina strains and their EPS to select indigenous bacteria able to grow in the presence of polycyclic aromatic hydrocarbons (naphthalene, phenanthrene and pyrene) from waste crude oil. The majority of isolated strains belonged to the genus Bacillus.

Moderately Halophilic, Exopolysaccharide-Producing Bacteria

Moderate halophiles include a wide array of microorganisms, taxonomically and physiologically distributed among many groups within the Bacteria domain and some groups of the Archaea. Their common characteristic is that they grow best at NaCl concentrations between 0.5 and 2.5 M (Kushner and Kamekura 1988), although they can be found in quite a diverse range of hypersaline habitats (Horikoshi and Grant 1998; Oren 1999).

Production of bioemulsifier by Bacillus subtilis, Alcaligenes faecalis and Enterobacter species in liquid culture

Three bacterial strains isolated from waste crude oil were selected due to their capacity of growing in the presence of hydrocarbons and production of bioemulsifier. The genetic identification (PCR of the 16S rDNA gene using fD1 and rD1 primers) of these strains showed their affiliation to Bacillus subtilis, Alcaligenes faecalis and Enterobacter sp. These strains were able to emulsify n-octane, toluene, xylene, mineral oils and crude oil, look promising for bioremediation application. Finally, chemical composition, emulsifying activity and surfactant activity of the biopolymers produced by the selected strains were studies under different culture conditions. Our results showed that chemical and functional properties of the bioemulsifiers were affected by the carbon source added to the growth media.

Some rheological properties of the extracellular polysaccharide produced byVolcaniella eurihalina F2-7

Volcaniella eurihalina strain F2-7 synthesizes an exopolysaccharide named V2-7, primarily composed of glucose, mannose, and rhamnose. The effect of chemical and physical factors on solution viscosity was studied. The V2-7 EPS showed pseudoplastic behavior at concentrations over 0.5% w/v. Viscosity decreased with temperature, but the viscosity values were restored after cooling. Freeze-thawing treatment did not affect the rheological properties of its solutions. Addition of inorganic salts produced a diminution of viscosity. However, the most remarkable aspect of V2-7 EPS is the effect of pH on its solutions; it is able to form high viscosity solutions, like a gel, at low pH values even in the presence of inorganic salts. This property, not present in neutral and alkaline solutions, makes it potentially useful for various industrial applications.

Effect of growth conditions on the rheological properties and chemical composition ofVolcaniella eurihalina exopolysaccharide

The exopolysaccharide produced byVolcaniella eurihalina, an halophilic eubacterium, under different environmental and nutritional conditions, is studied.V. eurihalina synthesizes an acidic heteropolysaccharide, composed by rhamnose, glucose, and mannose, as well as amino sugars, uronic acids, and acetyl and sulphate residues. This composition varies depending on the nutrients of culture medium. Viscosity and pseudoplasticity of the polymer solutions are also influenced by the nutritional conditions in which the microorganism was grown.