Lipopeptide production by Serratia marcescens SmSA using a Taguchi design and its application in enhanced heavy oil recovery.

Biosurfactant production at reactor level by Serratia marcescens SmSA was optimized and evaluated to enhance the heavy oil recovery on carbonate rocks. Temperature, agitation, and carbon/nitrogen (C/N) ratio were evaluated to optimize biosurfactant production by using a Taguchi (L9) design. The best conditions (C/N ratio: 6, 25 °C, and agitation: 100 rpm) were used to scale up the biosurfactant production with a 3-L bioreactor. The best aeration for biosurfactant production was 0.66 volume of air per volume of liquid per minute (vvm), producing the lowest surface tension (26 mN/m) in 14 h, with a biosurfactant yield of 14.26 g/L as a crude product and 2.85 g/L as a purified product, and a critical micelle concentration of 280 mg/L. The biosurfactant was characterized as a lipopeptide, and it was stable under extreme conditions: pH (2-12), salinity up to 200 g/L, and temperature up to 150 °C confirmed by thermogravimetric analysis. Enhanced oil recovery test was carried out with a carbonate core and heavy oil under reservoir conditions, obtaining an additional recovery of 8%, due to reduced interfacial tension and modified wettability of the rock. These findings highlight the potential application of S. marcescens SmSA biosurfactant in enhanced oil recovery.

Aerobic biodegradation of sludge with high hydrocarbon content generated by a Mexican natural gas processing facility.

The biodegradation of oil sludge from Mexican sour gas and petrochemical facilities contaminated with a high content of hydrocarbons, 334.7 ± 7.0 g kg(-1) dry matter (dm), was evaluated. Studies in microcosm systems were carried out in order to determine the capacity of the native microbiota in the sludge to reduce hydrocarbon levels under aerobic conditions. Different carbon/nitrogen/phosphorous (C/N/P) nutrient ratios were tested. The systems were incubated at 30 °C and shaken at 100 rpm. Hydrocarbon removals from 32 to 51% were achieved in the assays after 30 days of incubation. The best assay had C/N/P ratio of 100/1.74/0.5. The results of the Microtox(®) and Ames tests indicated that the original sludge was highly toxic and mutagenic, whereas the best assay gave a final product that did not show toxicity or mutagenicity.

Evaluation of haloalkaliphilic sulfur-oxidizing microorganisms with potential application in the effluent treatment of the petroleum industry.

Haloalkaliphilic sulfur-oxidizing mixed cultures for the treatment of alkaline-saline effluents containing sulfide were characterized and evaluated. The mixed cultures (IMP-PB, IMP-XO and IMP-TL) were obtained from Mexican alkaline soils collected in Puebla (PB), Xochimilco (XO) and Tlahuac (TL), respectively. The Ribosomal Intergenic Spacer Analysis (RISA) revealed bacteria related to Thioalkalibacterium and Thioalkalivibrio in IMP-XO and IMP-PB mixed cultures. Halomonas strains were detected in IMP-XO and IMP-TL. In addition, an uncultured Bacteroides bacterium was present in IMP-TL. Mixed cultures were evaluated at different pH and NaCl concentrations at 30°C. IMP-PB and IMP-TL expressed thiosulfate-oxidizing activity in the 7.5-10.5 pH range, whereas IMP-XO presented its maximal activity with 19.0 mg O2 g (protein)⁻¹ min⁻¹, at pH 10.6; it was not affected by NaCl concentrations up to 1.7 M. In continuous culture, IMP-XO showed a growth rate of 15 day⁻¹, productivity of 433.4 mg(protein) l⁻¹ day⁻¹ and haloalkaliphilic sulfur-oxidizing activity was also detected up to 170 mM by means of N-methyl-diethanolamine (MDEA). Saline-alkaline soil samples are potential sources of haloalkaliphilic sulfur-oxidizing bacteria and the mixed cultures could be applied in the treatment of inorganic sulfur compounds in petroleum industry effluents under alkaline-saline conditions.

Inhibition of the acetoclastic methanogenic activity by phenol and alkyl phenols.

Chemical and petrochemical industries are important sources of aromatic pollutants. Petrochemical processes like caustic washing of middle distillates produce the spent caustic liquors highly concentrated in phenol and alkyl phenols. The anaerobic technology is considered a feasible strategy for petrochemical wastewater pre-treatment although high concentrations of phenol could limit its efficiency. The goal of this work was to determine the toxicity of both selected alkyl phenols and a synthetic "spent-caustic phenols mixture" on the acetoclastic Specific Methanogenic Activity (SMA) of unadapted and phenol-adapted granular sludge. Alkyl phenols were responsible for 50% (IC50) and 100% (IC100) inhibition of the SMA at concentrations ranging from 1.6 to 5.0 mM and from 4.1 to 27.5 mM, respectively, for un-adapted granular sludge. In the case of phenol-adapted granular sludge, the inhibitory concentrations ranged from 1.7 to 14.9 mM and from 4.0 to 83.0 for IC50 and IC100, respectively, highlighting the impact of sludge acclimation. The inhibition produced by 2-ethylphenol was more acute compared to phenol and was not reduced by the phenol acclimation process. The IC50 and IC100 values obtained for the synthetic "spent-caustic phenols mixture" were 9.5 mM and 88.4 mM, respectively. The inhibitory concentrations of phenol compounds were closely correlated with compound apolarity (log P), indicating that the lipophilic character of the tested compounds was responsible for their methanogenic toxicity. An inhibition model is confirmed to estimate the IC50 and IC100.

Evaluation of the effect of nutrient ratios on biosurfactant production by Serratia marcescens using a Box-Behnken design.

The strain SmSA, identified as Serratia marcescens and known as a biosurfactant producer, was isolated from hydrocarbon contaminated soil from Veracruz, México. The interactions among the C/N, C/Mg and C/Fe ratios have not been examined for this microorganism. In this work was evaluated the effect of these nutrients at three levels using a mineral medium with glucose as the carbon source. A Box-Behnken experimental design was utilised to maximise biosurfactant production, which was assessed by oil spreading and surface tension tests. The treatment with C/N=5, C/Fe=26,000 and C/Mg=30 showed the best result since the surface tension was reduced to 30 mN m(-1). The multiple regression and response surface analyses indicated that the interaction between C/N and C/Mg had the utmost effect on the reduction of surface tension and biosurfactant production. The conditions of the best treatment were used to scale up biosurfactant production in a 3L bioreactor giving a yield of 4.1 gL(-1) of pure biosurfactant. It was found that the biosurfactant was mainly produced in the exponential phase and decreased the surface tension to 31 mN m(-1). The contact between the biosurfactant with heavy oil (15° API) increased its displacement from 9.3 to 18 cm.

Microcosm assays and Taguchi experimental design for treatment of oil sludge containing high concentration of hydrocarbons.

Microcosm assays and Taguchi experimental design was used to assess the biodegradation of an oil sludge produced by a gas processing unit. The study showed that the biodegradation of the sludge sample is feasible despite the high level of pollutants and complexity involved in the sludge. The physicochemical and microbiological characterization of the sludge revealed a high concentration of hydrocarbons (334,766+/-7001 mg kg(-1) dry matter, d.m.) containing a variety of compounds between 6 and 73 carbon atoms in their structure, whereas the concentration of Fe was 60,000 mg kg(-1) d.m. and 26,800 mg kg(-1) d.m. of sulfide. A Taguchi L(9) experimental design comprising 4 variables and 3 levels moisture, nitrogen source, surfactant concentration and oxidant agent was performed, proving that moisture and nitrogen source are the major variables that affect CO(2) production and total petroleum hydrocarbons (TPH) degradation. The best experimental treatment yielded a TPH removal of 56,092 mg kg(-1) d.m. The treatment was carried out under the following conditions: 70% moisture, no oxidant agent, 0.5% of surfactant and NH(4)Cl as nitrogen source.

Toxicity and kinetic parameters of the aerobic biodegradation of the phenol and alkylphenols by a mixed culture.

A mixed culture aerobically metabolized phenol, cresol isomers (o-,m-,p-), 2-ethylphenol and xylenol isomers (2,5-DMP and 3,4-DMP) as the sole carbon and energy source. This culture had a high tolerance towards phenol with values of maximum degradation rate (Vmax) of 47 microM phenol mg-1 protein h-1 and inhibition substrate constant (Ki) of 10 mM. These kinetic parameters were considerably diminished and the toxicity increased with the alkylphenols. For example with 2,5-xylenol, Vmax and Ki values of 0.8 microM 2,5-xylenol mg-1 protein h-1 and 1.3 mM, respectively, were obtained. The cresols were 5-fold more toxic than phenol, whereas 2-ethylphenol and 3,4-xylenol were 11-fold more toxic, and 2,5-xylenol was 34-fold more toxic than phenol.