Biosurfactants production by Pseudomonas aeruginosa FR using palm oil

Biosurfactants production by a strain of Pseudomonas aeruginosa using palm oil as a sole carbon source was investigated. The experiments were carried out in 500-mL conical flasks containing 100 mL of mineral media supplemented with palm oil as the sole carbon source. The P. aeruginosa FR strain was able to reduce surface tension of three tested inorganic media. Rotation velocities from 100 to 150 rpm provided free-cell fermented media with the lowest surface tension of approx 33 mN/m. Emulsification index results of even 100% were achieved when diesel was used as oil phase. Eight surface-active compounds produced by the bacterium were identified by mass spectrometry.     

Evaluation of different carbon and nitrogen sources in production of rhamnolipids by a strain of Pseudomonas aeruginosa

Culture conditions involving variations in carbon and nitrogen sources and different C:N ratios were examined with the aim of increasing productivity in the process of rhamnolipid synthesis by Pseudomonas aeruginosa. In addition to the differences in productivity, the use of different carbon sources resulted in several proportions related to the types of rhamnolipids synthesized (monorhamnolipids and dirhamnolipids). Furthermore, the variation in nutrients, mainly the nitrogen source, resulted in different amounts of virulence factors, as phenazines and extracellular proteins. The data point out a new concern in the choice of substrate to be used for rhamnolipid production by P. aeruginosa: toxic byproducts.     

Characterization by electrospray ionization and tandem mass spectrometry of rhamnolipids produced by two Pseudomonas aeruginosa strains isolated from Brazilian crude oil

In this work, biosurfactants produced by two Pseudomonas aeruginosa strains isolated from Brazilian crude oils were identified by proton nuclear magnetic resonance ((1)H NMR) and further characterized by mass spectrometry (MS) coupled with electrospray ionization (ESI) and tandem mass spectrometry (MS/MS) analysis in positive mode and their surface activities evaluated. Mono-rhamnolipids and di-rhamnolipids were identified for both isolates, but the most abundant were found to be mono-rhamnolipids. The similarity of rhamnolipids produced by the two strains was in good agreement with their surface activities. Both biosurfactants exhibited similar aqueous solution surface tensions, high emulsification indexes and critical micelle concentration values. The results obtained show that ESI-MS and MS/MS analysis alone provide a fast and highly specific characterization of biosurfactants produced by microbial strains.     

lux-marked Pseudomonas aeruginosa lipopolysaccharide production in the presence of rhamnolipid

Rhamnolipid produced by Pseudomonas aeruginosa during the late logarithmic growth phase has been reported to be an effective biosurfactant. In the presence of rhamnolipid, P. aeruginosa had a higher specific lipopolysaccharide production, which was attributed to the increase of carbon to nitrogen ratios owing to the increase of the solubility of hydrophobic hydrocarbons, favoring the synthesis of lipopolysaccharides. P. aeruginosa also had a higher ratio of lipopolysaccharides in the growth medium to those on cell surfaces than in the absence of rhamnolipid because the presence of rhamnolipid stimulated the release of lipopolysaccharides from P. aeruginosa cell outer membranes. The release of lipopolysaccharides from cell surfaces made P. aeruginosa exhibit a more hydrophobic surface, enhancing the accumulation of hydrophobic hydrocarbons on P. aeruginosa cell surfaces and consequently resulting in a higher growth rate.     

Microbial transformation of tributyltin chloride by Pseudomonas aeruginosa strain USS25 NCIM‐5224

A bacterial isolate capable of utilizing tributyltin chloride (TBTC) as sole source of carbon was isolated from marine water samples. The isolate, producing a soluble green pigment, was identified as Pseudomonas aeruginosa strain USS25 NCIM‐5224. The isolate showed maximum growth with 2 mM of TBTC in mineral salt medium. Time course results showed complete elimination of TBTC after 75 days of incubation. During the growth on TBTC, a product (280 mg) was found to accumulate, which was extracted with chloroform and detected on thin‐layer chromatography. Based on the IR, NMR spectra and GC‐MS analysis, the isolated product was identified as monobutyltin dichloro hydride. Copyright © 2005 John Wiley & Sons, Ltd.     

Microbial production of polyhydroxyalkanoates by bacteria isolated from oil wastes

A Gram-positive coccus-shaped bacterium capable of synthesizing higher relative molecular weight (M _r), polyhydroxybutyrate (PHB) was isolated from sesame oil and identified as Staphylococcus epidermidis (by Microbial ID, Inc., Newark, NJ). The experiment was conducted by shake flask fermentation culture using media containing fructose. Cell growth up to a dry mass of 2.5 g/L and PHB accumulation up to 15.02% of cell dry wt was observed. Apart from using single carbohydrate as a sole carbon source, various industrial food wastes including sesame oil, ice cream, malt, and soya wastes were investigated as nutrients for S. epidermidis to reduce the cost of the carbon source. As a result, we found that by using malt wastes as nutrient for cell growth, PHB accumulation of S. epidermidis was much better than using other wastes as nutrient source. The final dried cell mass and PHB production using malt wastes were 1.76 g/L and 6.93% polymer/cells (grams/gram), and 3.5 g/L and 3.31% polymer/cells (grams/gram) in shake flask culture and in fermentor culture, respectively. The bacterial polymer was characterized by ¹H-nuclear magnetic resonance (NMR), ¹³C-NMR, Fourier transform infrared, and differential scanning calorimetry. The results show that with different industrial food wastes as carbon and energy sources, the same biopolymer (PHB) was obtained. However, the use of sesame oil as the carbon source resulted in the accumulation of PHB with a higher melting point than that produced from other food wastes as carbon sources by this organism under similar experimental conditions.     

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.     

Siderophore production by a marine Pseudomonas aeruginosa and its antagonistic action against phytopathogenic fungi

A marine isolate of fluorescent Pseudomonas sp. having the ability to produce the pyoverdine type of siderophores under low iron stress (up to 10 μM iron in the succinate medium) was identified as Pseudomonas aeruginosa by using BIOLOG Breathprint and siderotyping. Pyoverdine production was optimum at 0.2% (w/v) succinate, pH 6.0, in an iron-deficient medium. Studies carried out in vitro revealed that purified siderophores and Pseudomonas culture have good antifungal activity against the plant deleterious fungi, namely, Aspergillus niger, Aspergillus flavus, Aspergillus oryzae, Fusarium oxysporum, and Sclerotium rolfsii. Siderophore-based maximum inhibition was observed against A. niger. These in vitro antagonistic actions of marine Pseudomonas against phytopathogens suggest the potential of the organism to serve as a biocontrol agent.     

Biodiesel fuel production by the transesterification reaction of soybean oil using immobilized lipase

The enzymatic alcoholysis of soybean oil with methanol and ethanol was investigated using a commercial, immobilized lipase (Lipozyme RMIM). The effect of alcohol (methanol or ethanol), enzyme concentration, molar ratio of alcohol to soybean oil, solvent, and temperature on biodiesel production was determined. The best conditions were obtained in a solvent-free system with ethanol/oil molar ratio of 3.0, temperature of 50 degrees C, and enzyme concentration of 7.0% (w/w). Three-step batch ethanolysis was most effective for the production of biodiesel. Ethyl esters yield was about 60% after 4 h of reaction.     

Optimization of alkaline transesterification of soybean oil and castor oil for biodiesel production

This article reports experimental data on the production of fatty acid ethyl esters from refined and degummed soybean oil and castor oil using NaOH as catalyst. The variables investigated were temperature (30–70°C), reaction time (1–3 h), catalyst concentration (0.5–1.5 w/wt%), and oil-to-ethanol molar ratio (1:3–1:9). The effects of process variables on the reaction conversion as well as the optimum experimental conditions are presented. The results show that conversions >95% were achieved for all systems investigated. In general, an increase in reaction temperature, reaction time, and in oil-to-ethanol molar ratio led to an enhancement in reaction conversion, whereas an opposite trend was verified with respect to catalyst concentration.     

Effects of rhamnolipid on degradation of granular organic substrate from kitchen waste by a Pseudomonas aeruginosa strain

The effect of rhamnolipid produced by a Pseudomonas aeruginosa strain on the aerobic degradation of granular organic substrate from kitchen waste by the bacterium was studied and compared with that of two synthetic surfactants, SDS and Triton X-100. The adsorption of rhamnolipid on the substrate, the surfactant-interfered adhesion of bacteria on the substrate as well as physicochemical and microbial conditions of the substrate during degradation were investigated. The adsorption isotherm of rhamnolipid on the substrate fit Freundlich law and its interactions with the substrate and bacteria weakened the adsorption of the bacteria on the substrate. The two synthetic surfactants, however, did not have such microbial effects. During degradation, rhamnolipid slowed down water evaporation in the substrate and significantly strengthened the dispersion of organic matter into the substrate water phase. The number of cells in the rhamnolipid treatment was higher than that in control and the remaining organic matter content in the substrates also had faster decreasing. SEM examination showed the on-site degradation of the substrate organic matter without rhamnolipid and the transfer of the degradation site in the presence of rhamnolipid. The results indicated that interference of rhamnolipid in the substrate matrix plays a potential role, physicochemically or microbially, on the degradation of the granular organic substrate. SDS and Triton X-100 may have the above physicochemical effects, but not so significant.     

Effects of plant lactones on the production of biofilm of Pseudomonas aeruginosa

Sixteen plant sesquiterpene lactones, thirteen from four species of the Family Asteraceae, and three from a species of Hepaticae, as well as seven annonaceous acetogenins isolated from the seeds of the tropical tree Annona cherimolia (Family Annonaceae), were evaluated for their ability to inhibit or stimulate the production of biofilm by a strain of Pseudomonas aeruginosa. The tested compounds carry a gamma-lactone moiety in their structures. This structural feature is similar to the lactone moiety present in N-acyl homoserine lactones, compounds that play the important role of "quorum sensors" in the mechanisms of biofilm formation observed in many gram-negative bacteria. A special assay was employed to evaluate the influence of the tested plant compounds to inhibit or stimulate the production of biofilm in a P. aeruginosa wild strain. Most of the tested compounds affected the biofilm formation mechanism. Six sesquiterpene lactones isolated from Acanthospermum hispidum and one from Enydra anagallis as well as an acetogenin from Annona cherimolia strongly inhibited (69-77%) the biofilm formation when incorporated to a bacterial culture at a concentration of 2.5 microg/ml. However, one of the acetogenins, squamocin, stimulated the biofilm formation even at a concentration of 0.25 microg/ml. The study of substances affecting the biofilm formation can lead to the design of new strategies to control P. aeruginosa infections.     

Effect of cations on the structure of bilayers formed by lipopolysaccharides isolated from Pseudomonas aeruginosa PAO1

The asymmetric outer membrane of Gram-negative bacteria contains lipopolysaccharides (LPSs) which contribute significantly to the bacterium's surface properties and play a crucial role in regulating membrane permeability. We report on neutron diffraction studies performed on aligned, self-assembled bilayers of Na-, Ca-, and Mg-salt forms of LPS isolated from Pseudomonas aeruginosa PAO1. From the one-dimensional neutron scattering length density profiles we find that water penetrates Ca2+-LPS bilayers to a lesser extent than either Na+- or Mg2+-LPS bilayers. This differential water penetration could have implications as to how small molecules permeate the outer membrane of Gram-negative bacteria and, possibly, how nonlamellar phases are formed.     

Production of biosurfactant from a new and promising strain of Pseudomonas aeruginosa PA1

The Pseudomonas aeruginosa PA1 strain, isolated from the water of oil production in Sergipe, Northeast Brazil, wasevaluated as a potential rhamnolipid type of biosurfactant producer. The production of biosurfactants was investigated using different carbon sources (n-hexadecane, paraffin oil, glycerol, and babassu oil) and inoculum concentrations (0.0016–0.008 g/L) The best results were obtained with glycerol as the substrate and an initial cell concentration of 0.004 g/L. AC:N ratio of 22.8 led to the greatest production of rhamnolipids (1700 mg/L) and efficiency (1.18 g of rhamnolipid/g of dry wt).     

Determination of residual oil in diesel oil by spectrofluorimetric and chemometric analysis

Multivariate calibration (PLS), principal components analysis (PCA) and linear discriminant analysis (LDA), associated to synchronous spectrofluorimetry, were used to identify and quantify non-transesterified residual vegetable oil in diesel oil with the addition of 2% of biodiesel (B2). The addition of residual oil, one of the easiest ways of adultering fuel, damages engines and leads to tax evasion. Using this method, the samples of diesel oil, B2, and B2 contaminated with residual oil were classified correctly and separated into three well-defined groups. The quantification of residual oil in B2 was carried out in the 0-25% (w/w) band, RMSEC and RMSEP values ranging from 0.26 to 0.48% (w/w) and 1.6-2.6% (w/w), respectively. The method is highly sensitive and efficient to identify and quantify this type of adulterant in which 100% of the samples were correctly classified and the average relative error was approximately 4% in the range 0.5-25% (w/w).     

Photochemical inactivation of Pseudomonas aeruginosa

Adaptability to a broad range of environments together with relatively high resistance to antibiotics and to disinfectants makes Pseudomonas aeruginosa a concern in hospitals and in public health. We investigated whether UVA-mediated photochemical inactivation of P. aeruginosa could be accomplished with high efficiency while at the same time preserving the sensitivity of subsequent diagnostic tests. We characterized dose responses and bactericidal kinetic rates of 5-iodonaphthyl 1-azide (INA) and of amotosalen (AMO) as these substances exposed to UVA are known to inactivate germs with minimal impact to blood products or to viral antigens. Neither UVA without photochemicals nor INA or AMO in the dark inactivated bacteria. We found that AMO was ca 1000-fold more effective in inactivating P. aeruginosa cells than INA under similar conditions. Photoinactivation with either INA or AMO at conditions that abolished bacterial infectivity did not impair polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) testing. For comparison, similar titers of Bacillus atrophaeus spores (a surrogate for B. anthracis) remained unaffected at conditions that reduced the survival of P. aeruginosa below detection levels. The results presented in this study should assist in improved methods to inactivate P. aeruginosa in environmental, clinical and forensic samples without impairing subsequent nucleic acid- or immune-based analysis.     

Multivariate monitoring of soybean oil ethanolysis by FTIR

In this work, an analytical procedure was developed to monitor the ethanolysis of degummed soybean oil (DSO) using Fourier-transformed mid-infrared spectroscopy (FTIR) and methods of multivariate analysis such as principal component analysis (PCA) and partial least squares regression (PLS). The triglycerides (reagents) and ethyl esters (products) involved in ethanolysis were shown to have similar FTIR spectra. However, when the FTIR spectra derived from seven standard mixtures of triolein and ethyl oleate were treated by PCA at the region that represents the CO stretching vibration of ester groups (1700-1800 cm⁻¹), only two principal components (PC) were shown to capture 99.95% of the total spectral variance (92.37% for the former and 7.58% for the latter PC). This observation supported the development of a multivariate calibration model that was based on the PLS regression of the FTIR data. The prevision capability of this model was measured against 40 reaction aliquots whose ester content was previously determined by size exclusion chromatography. Only small discrepancies were observed when the two experimental data sets were treated by linear regression (R²=0.9837) and these deviations were attributed to the occurrence of non-modeled transient species in the reaction mixture (reaction intermediates), particularly at short reaction times. Therefore, the FTIR/PLS model was shown to be a fast and accurate method to predict reaction yields and to follow the in situ kinetics of soybean oil ethanolysis.