The influence of vegetable oils on biosurfactant production by Serratia marcescens

The production of biosurfactant, a surface-active compound, by two Serratia marcescens strains was tested on minimal culture medium supplemented with vegetable oils, considering that it is well known that these compounds stimulate biosurfactant production. The vegetable oils tested included soybean, olive, castor, sunflower, and coconut fat. The results showed a decrease in surface tension of the culture medium without oil from 64.54 to 29.57, with a critical micelle dilution (CMD⁻¹) and CMD⁻² of 41.77 and 68.92 mN/m, respectively. Sunflower oil gave the best results (29.75 mN/m) with a CMD⁻¹ and CMD⁻² of 36.69 and 51.41 mN/m, respectively. Sunflower oil contains about 60% of linoleic acid. The addition of linoleic acid decreased the surface tension from 53.70 to 28.39, with a CMD⁻¹ of 29.72 and CMD⁻² of 37.97, suggesting that this fatty acid stimulates the biosurfactant production by the LB006 strain. In addition, the crude precipitate surfactant reduced the surface tension of water from 72.00 to 28.70 mN/m. These results suggest that the sunflower oil’s linoleic acid was responsible for the increase in biosurfactant production by the LB006 strain.     

Biosurfactant production under extreme environmental conditions by an efficient microbial consortium, ERCPPI-2

The biosurfactant production potential of a new microbial consortium of Enterobacter cloacae and Pseudomonas sp. (ERCPPI-2) which was isolated from heavy crude oil-contaminated soil in the south of Iran, has been investigated under extreme environmental conditions. The isolated consortium produces a biosurfactant mixture with excessive oil spreading and emulsification properties. This consortium was able to grow and produce biosurfactant at temperatures up to 70 °C, pressures up to 6000 psia, salinities up to 15% (w/v), and in the pH range 4-10. Besides, the optimum biosurfactant production conditions were found to be 40 °C and 7.0 for the temperature and pH value, respectively. These conditions gave the best biosurfactant production of 1.74 g/1 when the cells were grown on a minimal salt medium containing 1.0% (w/v) olive oil, 1.0% (w/v) sodium nitrate supplemented with 1.39% (w/v) K(2)HPO(4) at 40 °C and 150 rpm after 48 h of incubation. The ERCPPI-2 could reduce surface and interfacial tensions to 31.7 and 0.65 mN/m from the original values of 58.3 and 16.9 mN/m, respectively. The isolated consortium produced biosurfactant using heavy crude oil as the sole source of carbon and emulsified the available heavy crude oil up to E(24)=83.4%. The results of the core holder flooding tests at simulated reservoir conditions demonstrated that the oil recovery efficiency due to the injection of the cell-free biosurfactant solution was 27.2%, and the bacterium injection reduced the final residual oil saturations to below 3% at optimum conditions.     

Production and characterization of biosurfactant from marine Streptomyces species B3

The present study demonstrates the production and properties of a biosurfactant isolated from marine Streptomyces species B3. The production of the biosurfactant was found to be higher in medium containing sucrose and lower in the medium containing glycerol. Yeast extract was the best nitrogen source for the production of the biosurfactant. The isolated biosurfactant reduced the surface tension of water to 29 mN/m. The purified biosurfactant was shown critical micelle concentrations of 110 mg/l. The emulsifying activity and stability of the biosurfactant was investigated at different salinities, pH, and temperature. The biosurfactant was effective at very low concentrations over a wide range of temperature, pH, and salt concentration. The purified biosurfactant was shown strong antimicrobial activity. The biosurfactant was produced from the marine Streptomyces sp. using non-hydrocarbon substrates such as sucrose that was readily available and not required extensive purification procedure. Streptomyces species B3 can be used for microbially enhanced oil recovery process.     

Utilization of Banana Peel as a Novel Substrate for Biosurfactant Production by Halobacteriaceae archaeon AS65

In this study, biosurfactant-producing bacteria was evaluated for biosurfactant production by using banana peel as a sole carbon source. From the 71 strains screened, Halobacteriaceae archaeon AS65 produced the highest biosurfactant activity. The highest biosurfactant production (5.30 g/l) was obtained when the cells were grown on a minimal salt medium containing 35 % (w/v) banana peel and 1 g/l commercial monosodium glutamate at 30 °C and 200 rpm after 54 h of cultivation. The biosurfactant obtained by extraction with ethyl acetate showed high surface tension reduction (25.5 mN/m), a small critical micelle concentration value (10 mg/l), thermal and pH stability with respect to surface tension reduction and emulsification activity, and a high level of salt tolerance. The biosurfactant obtained was confirmed as a lipopeptide by using a biochemical test FT-IR, NMR, and mass spectrometry. The crude biosurfactant showed a broad spectrum of antimicrobial activity and had the ability to emulsify oil, enhance PAHs solubility, and oil bioremediation.     

First Report of a Lipopeptide Biosurfactant from Thermophilic Bacterium Aneurinibacillus thermoaerophilus MK01 Newly Isolated from Municipal Landfill Site

A biosurfactant-producing thermophile was isolated from the Kahrizak landfill of Tehran and identified as a bacterium belonging to the genus Aneurinibacillus. A thermostable lipopeptide-type biosurfactant was purified from the culture medium of this bacterium and showed stability in the temperature range of 20–90 °C and pH range of 5–10. The produced biosurfactant could reduce the surface tension of water from 72 to 43 mN/m with a CMC of 1.21 mg/mL. The strain growing at a temperature of 45 °C produces a substantial amount of 5 g/L of biosurfactant in the medium supplemented with sunflower oil as the sole carbon source. Response surface methodology was employed to optimize the biosurfactant production using sunflower oil, sodium nitrate, and yeast extract as variables. The optimization resulted in 6.75 g/L biosurfactant production, i.e., 35 % improved as compared to the unoptimized condition. Thin-layer chromatography, FTIR spectroscopy, ¹H-NMR spectroscopy, and biochemical composition analysis confirmed the lipopeptide structure of the biosurfactant.     

Lipopeptide surfactant production by Bacillus subtilis grown on low-cost raw materials

The production of biosurfactant by Bacillus subtilis ATCC 6633 was investigated using commercial sugar, sugarcane juice and cane molasses, sugarcane juice alcohol stillage, glycerol, mannitol, and soybean oil. Commercial sugar generated the minimum values of surface tension, with the best results (28.7 mN/m, (relative critical micelle concentration [CMC⁻¹] of 78.6) being achieved with 10 g of substrate/L in 48 h. At a pH between 7.0 and 8.0, a higher production of surface-active compounds and a greater emulsifier activity was also observed. Enrichment of the culture medium with trace minerals and EDTA showed maximum yields, whereas supplementation with yeast extract stimulated only cell growth. The kinetic studies revealed that biosurfactant production is a cell growth-associated process; surface tension, CMC, and emulsification index values of 29.6 dyn/cm, 82.3, and 57%, respectively, were achieved, thus indicating that it is feasible to produce biosurfactants from a renewable and low-cost carbon source.     

Production and properties of a biosurfactant obtained from a member of the Bacillus subtilis group (PTCC 1696)

The production and properties of a biosurfactant, synthesized by a member of the Bacillus subtilis group (PTCC 1696) which was isolated from an Iranian oil field, have been investigated. The biosurfactant, which was produced as a primary metabolite associated with the growth of PTCC 1696, was able to reduce the surface and interfacial tension of media to 26.7 and 0.1 mN/m, respectively. Crude biosurfactant and acid precipitated biosurfactant have critical micelle concentrations of 10 and 100 mug/ml, respectively. The stability of the biosurfactant at different salinities, pH and temperature and also its emulsifying activity have been investigated. It is an effective surfactant at very low concentrations over a wide range of temperatures, pHs and salt concentrations and also has the ability to emulsify oil, which is essential for enhanced oil recovery.     

Analysis of Rhamnolipid Biosurfactants Produced Through Submerged Fermentation Using Orange Fruit Peelings as Sole Carbon Source

The fermentative production of rhamnolipid biosurfactant from Pseudomonas aeruginosa MTCC 2297 was carried out by submerged fermentation using various cost-effective waste materials such as orange peelings, carrot peel waste, lime peelings, coconut oil cake, and banana waste. The orange peel was found to be the best substrate generating 9.18 g/l of rhamnolipid biosurfactant with a surface tension reduction up to 31.3 mN/m. The production was growth independent, and optimum conditions were standardized. The emulsifying activity was highest against kerosene (73.3%). Rhamnolipid components were purified and separated by ethyl acetate extraction, preparative silica gel column chromatography, high-performance liquid chromatography and thin-layer chromatography. The major rhamnolipid components were characterized, by fast atom bombardment mass spectrometry, as a mixture of dirhamnolipids and monorhamnolipids.     

Interactive optimization of biosurfactant production by Paenibacillus alvei ARN63 isolated from an Iranian oil well

The potential of an indigenous bacterial strain isolated from an Iranian oil field for the production of biosurfactant was investigated in this study. After isolation, the bacterium was characterized to be Paenibacillus alvei by biochemical tests and 16S ribotyping. The biosurfactant, which was produced by this bacterium, was able to lower the surface tension of media to 35 mN/m. Accordingly, thin layer chromatography (TLC) and FT-IR has been carried out to determine compositional analysis of the produced biosurfactant. After all the tests related to characterization of the biosurfactant produced by the isolated bacterium, it was characterized as lipopeptide derivative. The combination of central composite rotatable design (CCRD) and response surface methodology (RSM) was exploited to optimize biosurfactant production. Therefore, variations of four impressive parameters, pH, temperature, glucose and salinity concentrations were selected for optimization of growth conditions. The empirical model developed through RSM in terms of effective operational factors mentioned above was found to be adequate to describe the biosurfactant production. A maximum reduction in surface tension was obtained under the optimal conditions of 13.03 g/l glucose concentration, 34.76 °C, 51.39 g/l total salt concentration and medium pH 6.89.     

Biosurfactant production by Bacillus subtilis using cassava-processing effluent

A cassava flour-processing effluent (manipueira) was evaluated as a substrate for surfactant production by two Bacillus subtilis strains. B. subtilis ATCC 21332 reduced the surface tension of the medium to 25.9 mN/m, producing a crude biosurfactant concentration of 2.2 g/L. The wild-type strain, B. subtilis LB5a, reduced the surface tension of the medium to 26.6 mN/m, giving a crude biosurfactant concentration of 3.0 g/L. A decrease in surfactant concentration observed for B. subtilis ATCC 21332 seemed to be related to an increase in protease activity. The biosurfactant produced on cassava effluent medium by B. subtilis LB5a was similar to surfactin.     

Biosurfactant production by Rhodococcus erythropolis grown on glycerol as sole carbon source

The production of biosurfactant by Rhodococcus erythropolis during the growth on glycerol was investigated. The process was carried out at 28 degrees C in a 1.5-L bioreactor using glycerol as carbon source. The bioprocess was monitored through measurements of biosurfactant concentration and glycerol consumption. After 51 h of cultivation, 1.7 g/L of biosurfactant, surface, and interfacial tensions values (with n-hexadecane) of 43 and 15 mN/m, respectively, 67% of Emulsifying Index (E (24)), and 94% of oil removal were obtained. The use of glycerol rather than what happens with hydrophobic carbon source allowed the release of the biosurfactant, originally associated to the cell wall.     

Synthesis,Characterization, and Oil Recovery Application of Biosurfactant Produced by Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> WJ-1 Using Waste Vegetable Oils

A bacterial strain was isolated and cultured from the oil excavation areas in tropical zone in northern China. The biochemical characteristics and partial sequenced 16S rRNA gene of isolate, WJ-1, was identical to those of cultured representatives of the species Pseudomonas aeruginosa. This bacterium was able to produce a type of biosurfactant. Compositional analysis revealed that the extracted biosurfactant was composed of high percentage lipid (∼74%, w/w) and carbohydrate (∼20%, w/w) in addition to a minor fraction of protein (∼6%, w/w). The best production of 50.2 g/l was obtained when the cells were grown on minimal salt medium containing 6.0% (w/v) glucose and 0.75% (w/v) sodium nitrate supplemented with 0.1% (v/v) element solution at 37 °C and 180 rpm after 96 h. The optimum biosurfactant production pH value was found to be 6.0–8.0. The biosurfactant of WJ-1, with the critical micelle concentration of 0.014 g/L, could reduce surface tension to 24.5 mN/m and emulsified kerosene up to EI₂₄ ≈95. The results obtained from time course study indicated that the surface tension reduction and emulsification potential was increased in the same way to cell growth. However, maximum biosurfactant production occurred and established in the stationary growth phase (after 90 h). Thin layer chromatography, Fourier transform infrared spectrum, and mass spectrum analysis indicate the extracted biosurfactant was affiliated with rhamnolipid. The core holder flooding experiments demonstrated that the oil recovery efficiency of strain and its biosurfactant was 23.02% residual oil.     

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.     

Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir

Biosurfactant produced by Pseudomonas aeruginosa, Bacillus subtilis and Rhodococcus erythropolis that isolated from the formation water of Chinese petroleum reservoir has been compared in surface abilities and oil recovery. Maximum biosurfactant production reached to about 2.66 g/l and the surface tension of liquid decreased from 71.2 to 22.56 mN/m using P. aeruginosa. Three strains exhibited a good ability to emulsify the crude oil, and biosurfactant of P. aeruginosa attained an emulsion index of 80% for crude oil which was greater than other strains. Stability studies were carried out under the extreme environmental conditions, such as high temperature, pH, salinity and metal ions. Results showed an excellent resistance of all biosurfactants to retain their surface-active properties at extreme conditions. It was found that the biosurfactants from three isolated bacteria showed a good stability above pH of 5, but at lower pH (from 1 to 5) they will harmfully be affected. They were able to support the condition up to 20 g/l salinity. P. aeruginosa biosurfactant was even stable at the higher salinity. Regarding temperature, all produced biosurfactants demonstrated a good stability in the temperature up to 120 °C. But stability of three biosurfactants was affected by monovalent and trivalent ions. Oil recovery experiments in physical simulation showed 7.2-14.3% recovery of residual oil after water flooding when the biosurfactant of three strains was added. These results suggest that biosurfactants of these indigenous isolated strains are appropriate candidates for enhanced oil recovery with a preference to biosurfactant of P. aeruginosa.     

Microbial surfactant-enhanced mineral oil recovery under laboratory conditions

Microbial enhanced oil recovery (MEOR) is potentially useful to recover incremental oil from a reservoir being beyond primary and secondary recovery operations. Effort has been made to isolate and characterize natural biosurfactant produced by bacterial isolates collected from various oil fields of ONGC in Assam. Production of biosurfactant has been considered to be an effective major index for the purpose of enhanced oil recovery. On the basis of the index, four promising bacterial isolates: Pseudomonas aeruginosa (MTCC7815), P. aeruginosa (MTCC7814), P. aeruginosa (MTCC7812) and P. aeruginosa (MTCC8165) were selected for subsequent testing. Biosurfactant produced by the promising bacterial isolates have been found to be effective in the recovery of crude oil from saturated column under laboratory conditions. Two bacterial strains: P. aeruginosa (MTCC7815) and P. aeruginosa (MTCC7812) have been found to be the highest producer of biosurfactant. Tensiometer studies revealed that biosurfactants produced by these bacterial strains could reduce the surface tension (sigma) of the growth medium from 68 to 30 mN m(-1) after 96 h of growth. The bacterial biosurfactants were found to be functionally stable at varying pH (2.5-11) conditions and temperature of 100 degrees C. The treatment of biosurfactant containing, cell free culture broth in crude oil saturated sand pack column could release about 15% more crude oil at 90 degrees C than at room temperature and 10% more than at 70 degrees C under laboratory condition.     

Biosurfactant production by Rhodococcus erythropolis grown on glycerol as sole carbon source

The production of biosurfactant by Rhodococcus erythropolis during the growth on glycerol was investigated. The process was carried out at 28°C in a 1.5-L bioreactor using glycerol as carbon source. The bioprocess was monitored through measurements of biosurfactant concentration and glycerol consumption. After 51 h of cultivation, 1.7 g/L of biosurfactant, surface, and interfacial tensions values (with n-hexadecane) of 43 and 15 mN/m, respectively, 67% of Emulsifying Index (E ₂₄), and 94% of oil removal were obtained. The use of glycerol rather than what happens with hydrophobic carbon source allowed the release of the biosurfactant, originally associated to the cell wall.     

An efficient biosurfactant-producing bacterium Pseudomonas aeruginosa MR01, isolated from oil excavation areas in south of Iran

A bacterial strain was isolated and cultured from the oil excavation areas in tropical zone in southern Iran. It was affiliated with Pseudomonas. The biochemical characteristics and partial sequenced 16S rRNA gene of isolate, MR01, was identical to those of cultured representatives of the species Pseudomonas aeruginosa. This bacterium was able to produce a type of biosurfactant with excessive foam-forming properties. Compositional analysis revealed that the extracted biosurfactant was composed of high percentages lipid (65%, w/w) and carbohydrate (30%, w/w) in addition to a minor fraction of protein (4%, w/w). The best production of 2.1 g/l was obtained when the cells were grown on minimal salt medium containing 1.2% (w/v) glucose and 0.1% (w/v) ammonium sulfate supplemented with 0.1% (w/v) isoleucine at 37 °C and 180 rpm after 2 days. The optimum biosurfactant production pH value was found to be 8.0. The MR01 could reduce surface tension to 28 mN/m and emulsified hexadecane up to E24  70. The results obtained from time course study indicated that the surface tension reduction and emulsification potential was increased in the same way to cell growth. However, maximum biosurfactant production occurred and established in the stationary growth phase (after 84 h). Fourier Transform Infrared spectrum of extracted biosurfactant indicates the presence of carboxyl, amine, hydroxyl and methoxyl functional groups. Thermogram of biosurfactant demonstrated three sharp endothermic peaks placing between 200 and 280 °C. The core holder flooding experiments demonstrated that the oil recovery efficiencies varied from 23.7% to 27.1% of residual oil.     

STEROID-SAPONINS FROM FENUGREEK SEEDS: EXTRACTION,PURIFICATION AND SURFACE PROPERTIES

Two techniques for extraction and purification were excersized in order to obtain a purified saponins extract from several sources (fenugreek, soybean, locust bean and sesame). The results indicate that fenugreek steroid saponins extract has the best surface properties and reduces the surface tension to 33.3 mN/m and the tetradecane-water interfacial tension from 44 mN/m to 7.5 mN/m. The HLB value was evaluated and found to be 18. Three different types of oil (n-tetradecane, soybean oil, tricaprylin) were emulsified with a 6 wt% aqueous solution of saponins to give an average droplet size of 0.7 μm (yet stable after month). An effective synergism is seen when these emulsions were prepared with sucrose fatty acid esters as co-surfactant. Furthermore, the foaming power was evaluated.     

Cassava flour wastewater as a substrate for biosurfactant production

Five cassava flour wastewater (manipueira) preparations were tested as culture media for biosurfactant production by a wild-type Bacillus sp. isolate. No-solids (F), no-solids diluted (F/2), natural (I), natural diluted (I/2), and decanted (IPS) were the tested manipueira media. The microorganism was able to grow and to produce biosurfactant on all manipueira preparations. The media whose solids were removed (F and F/2) showed better results than preparations with the presence of solids (I, I/2, and IPS). No-solids medium (F) showed a surface tension of 26,59 mN/m and reciprocal of critical micelle concentration of over 100 and was selected as a potential substrate for biosurfactant production.     

Effect of boron on the surface tension of molten silicon and its temperature coefficient

The influence of boron concentration (C(B)/mass%) on the surface tension of molten silicon has been investigated with the sessile drop method under oxygen partial pressure P(O(2))=1.62x10(-25)-2.63x10(-22) MPa, and the results can be summarized as follows. The surface tension increases with C(B) in the range below 2.09 mass%, and the maximum increase rate of the surface tension is about 30 mN m(-1)(mass% C(B))(-1). The temperature coefficient of the surface tension, ( partial differential sigma/ partial differential T)C(B), was found to increase with the boron concentration in molten silicon. At the interface between molten silicon and the BN substrate, a discontinuous Si(3)N(4) layer was reckoned to form and the layer might prevent BN from dissolving into the molten silicon. Since dissolved boron from the BN substrate into the molten silicon is below 0.054 mass% and the associated increase in surface tension is below 1.5 mN m(-1), the contamination from the BN substrate on the surface tension can be ignored. The relation between the surface tension and C(B) indicates negative adsorption of boron and can be well described by combining the Gibbs adsorption isotherm with the Langmuir isotherm.