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 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.

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.

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.

A field trial for an ex-situ bioremediation of a drilling mud-polluted site.

The remediation of drilling mud-polluted sites in the Southeast of Mexico is a top priority for Mexican oil industry. The objective of this work was to find a technology to remediate these sites. A field trial was performed by composting in biopiles, where four 1ton soil-biopiles were established, one treatment in triplicate and one unamended biopile. Amended biopiles were added with nutrients to get a C/N/P ratio of 100/3/0.5 plus a bulking agent (straw) at a soil/straw ratio of 97/3. Moisture content was maintained around 30-35%. Results showed that, after 180 d, total petroleum hydrocarbon (TPH) concentrations decreased from 99300+/-23000mgTPHkg(-1) soil to 5500+/-770mgTPHkg(-1) for amended biopiles and to 22900+/-7800mgTPHkg(-1) for unamended biopile. An undisturbed soil control showed no change in TPH concentrations. Gas chromatographic analysis showed residual alkyl dibenzothiophene type compounds. Highest bacterial counts were observed during the first 30 d which correlated with highest TPH removal, whereas fungal count increased at the end of the experimentation period. Results suggested an important role of the straw, nutrient addition and water content in stimulating aerobic microbial activity and thus hydrocarbon removal. This finding opens an opportunity to remediate old polluted sites with recalcitrant and high TPH concentration.

Starch-based plastic polymer degradation by the white rot fungus Phanerochaete chrysosporium grown on sugarcane bagasse pith: enzyme production.

In this study, starch metabolites and enzymes were determined during starch-based plastic polymer biodegradation by the white rot fungus Phanerochaete chrysosporium, grown in sugarcane bagasse pith in tubular reactors. Various metabolites, amylase, ligninase and cellulase production were measured during P. chrysosporium growth on sugarcane bagasse pith with added glucose and starch polymer. On-line respirometric analyses followed during 32 days confirmed the P. chrysosporium capability of growing on sugarcane bagasse pith with starch polymer degradation. Enzyme activity during secondary metabolism increased, and a 70% and 74% starch degradation was reached with and without glucose addition, generating low molecular weight metabolites (e.g.) dextrin, maltotriose, maltose and glucose that were detected by high performance liquid chromatography.