Two schemes for production of biosurfactant from Pseudomonas aeruginosa MR01: Applying residues from soybean oil industry and silica sol-gel immobilized cells.

Rhamnolipids are the most common biosurfactants and P. aeruginosa strains are the most frequently studied microorganisms for the production of rhamnolipids. Eco-friendly advantages and promising applications of rhamnolipids in various industries are the major reasons for pursuing the economic production of these biosurfactants. This study shows that cultivation of P. aeruginosa MR01 in medium contained inexpensive soybean oil refinery wastes which exhibited similar levels and homologues of rhamnolipids. Mass spectrometry indicated that the Rha-C10-C10 and Rha-Rha-C10-C10 constitute the main rhamnolipids in different cultures of MR01 including one of oil carbon source analogues. Moreover, rhamnolipid mixtures extracted from different cultures showed critical micelle concentrations (CMC) in the range of ≃24 to ≃36mg/l with capability to reduce the surface tension of aqueous solution from 72 to ≃27-32mN/m. However, the sol-gel technique using tetraethyl orthosilicate (TEOS) was used as a gentler method in order to entrap the P. aeruginosa MR01 cells in mold silica gels. Immobilized cells can be utilized several times in consecutive fermentation batches as well as in flow fermentation processes. In this way, reusability of the cells may lead to a more economical fermentation process. Approximately 90% of cell viability was retained during the silica sol-gel immobilization and ≃84% of viability of immobilized cells was preserved for 365days of immobilization and storage of the cells in phosphate buffer at 4°C and 25°C. Moreover, mold gels showed good mechanical stability during the seven successive fermentation batches and the entrapped cells were able to efficiently preserve their biosurfactant-producing potential.

Management of soybean oil refinery wastes through recycling them for producing biosurfactant using Pseudomonas aeruginosa MR01.

Biosurfactant production through a fermentation process involving the biodegradation of soybean oil refining wastes was studied. Pseudomonas aeruginosa MR01 was able to produce extracellular biosurfactant when it was cultured in three soybean oil refinement wastes; acid oil, deodorizer distillate and soapstock, at different carbon to nitrogen ratios. Subsequent fermentation kinetics in the three types of waste culture were also investigated and compared with kinetic behavior in soybean oil medium. Biodegradation of wastes, biosurfactant production, biomass growth, nitrate consumption and the number of colony forming units were detected in four proposed media, at specified time intervals. Unexpectedly, wastes could stimulate the biodegradation activity of MR01 bacterial cells and thus biosurfactant synthesis beyond that of the refined soybean oil. This is evident from higher yields of biodegradation and production, as revealed in the waste cultures (Ydeg|(Soybean oil) = 53.9 % < Ydeg|(wastes) and YP/S|(wastes) > YP/S|(Soybean oil) = 0.31 g g(-1), respectively). Although production yields were approximately the same in the three waste cultures (YP/S|(wastes) =/~ 0.5 g g(-1)), microbial activity resulted in higher yields of biodegradation (96.5 ± 1.13 %), maximum specific growth rate (µ max = 0.26 ± 0.02 h(-1)), and biosurfactant purity (89.6 %) with a productivity of 14.55 ± 1.10 g l(-1), during the bioconversion of soapstock into biosurfactant. Consequently, applying soybean oil soapstock as a substrate for the production of biosurfactant with commercial value has the potential to provide a combination of economical production with environmental protection through the biosynthesis of an environmentally friendly (green) compound and reduction of waste load entering the environment. Moreover, this work inferred spectrophotometry as an easy method to detect rhamnolipids in the biosurfactant products.

Structural characterization of a rhamnolipid-type biosurfactant produced by Pseudomonas aeruginosa MR01: enhancement of di-rhamnolipid proportion using gamma irradiation.

We previously reported that MR01, an indigenous strain of Pseudomonas aeruginosa, was able to produce a rhamnolipid-type biosurfactant. Here, we attempted to define the structural properties of this natural product. The analysis of the extracted biosurfactant by thin-layer chromatography (TLC) revealed the presence of two compounds corresponding to those of authentic mono- and di-rhamnolipid. The identity of two structurally distinguished rhamnolipids was confirmed by 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. Liquid chromatography/mass spectrometry (LC/MS) of extracted biosurfactant revealed up to seventeen different rhamnolipid congeners. Further quantification showed di-rhamnolipids as the major compound (77.2%), while monorhamnolipids comprising a smaller proportion (22.8%) of MR01 biosurfactant. Rha-Rha-C10-C10 was verified as the major component of the MR01 biosurfactant (35.93%). Cytotoxic activity of MR01 biosurfactant against human cancer Hela cells showed an excellent inhibitory effect of 5µg/ml. An isolated mutant strain (MR01-C) created by Gamma ray irradiation demonstrated more than one and a half-fold biosurfactant production and activity compared with the parent strain. Analysis of the biosurfactant produced by MR01-C showed the magnitude of di-rhamnolipids in the sample increased up to 88.6% (∼15% higher than control) and the quantity of Rha-Rha-C10-C10 increased to 52.08% (∼45% higher than control).

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 ( approximately 65%, w/w) and carbohydrate ( approximately 30%, w/w) in addition to a minor fraction of protein ( approximately 4%, w/w). The best production of 2.1g/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 degrees C and 180rpm after 2 days. The optimum biosurfactant production pH value was found to be 8.0. The MR01 could reduce surface tension to 28mN/m and emulsified hexadecane up to E24 approximately 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 84h). 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 degrees C. The core holder flooding experiments demonstrated that the oil recovery efficiencies varied from 23.7% to 27.1% of residual oil.

Effective factors in the treatment of kerosene-water emulsion by using UF membranes.

The effects of different parameters including membrane type (regenerated cellulose and polysulphone), transmembrane pressure (TMP), the content of oil in the feed, the flow velocity of the feed and pH on the ultrafiltration of an emulsion of kerosene in water were studied. It was found that the important factors affecting ultrafiltration were, in order, membrane type, pressure and oil concentration. The greatest flux at the optimum conditions here of 3 bar, an oil content of 3% (v/v) and with membrane type C30F was predicted as 108 L/(m(2)h) that was within the range of the confidence limit of the measured value of 106 L/(m(2)h). The normalised FTIR results of the virgin cellulosic membranes C30F and C100F showed more abundant OH groups. The bigger number of OH groups implies a greater hydrophilicity. The larger observed flux in the C30F is related to a higher number of pores as well (surface porosity) compared with the C100F membrane. In the "polarised regime" from 3 bar upwards, flux was independent of pressure for all membranes and was assumed to be determined by the back diffusion transport. Despite the fact that both the PS100H and C100F membranes had the same cut-off (100 kg/mol), the hydrophilic C100F showed a superior permeate flux. The strongest drop of flux with time due to oil fouling was observed for the C100F although it was hydrophilic. In the case of the PS100H, both FTIR and SEM showed that cake layer formation was not the cause of fouling. Meanwhile the SEM and FTIR results of fouled C100F provided evidence of adsorptive and gel formation fouling.