19 research outputs found
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Development of an In Situ Biosurfactant Production Technology for Enhanced Oil Recovery
The long-term economic potential for enhanced oil recovery (EOR) is large with more than 300 billion barrels of oil remaining in domestic reservoirs after conventional technologies reach their economic limit. Actual EOR production in the United States has never been very large, less than 10% of the total U. S. production even though a number of economic incentives have been used to stimulate the development and application of EOR processes. The U.S. DOE Reservoir Data Base contains more than 600 reservoirs with over 12 billion barrels of unrecoverable oil that are potential targets for microbially enhanced oil recovery (MEOR). If MEOR could be successfully applied to reduce the residual oil saturation by 10% in a quarter of these reservoirs, more than 300 million barrels of oil could be added to the U.S. oil reserve. This would stimulate oil production from domestic reservoirs and reduce our nation's dependence on foreign imports. Laboratory studies have shown that detergent-like molecules called biosurfactants, which are produced by microorganisms, are very effective in mobilizing entrapped oil from model test systems. The biosurfactants are effective at very low concentrations. Given the promising laboratory results, it is important to determine the efficacy of using biosurfactants in actual field applications. The goal of this project is to move biosurfactant-mediated oil recovery from laboratory investigations to actual field applications. In order to meet this goal, several important questions must be answered. First, it is critical to know whether biosurfactant-producing microbes are present in oil formations. If they are present, then it will be important to know whether a nutrient regime can be devised to stimulate their growth and activity in the reservoir. If biosurfactant producers are not present, then a suitable strain must be obtained that can be injected into oil reservoirs. We were successful in answering all three questions. The specific objectives of the project were (1) to determine the prevalence of biosurfactant producers in oil reservoirs, and (2) to develop a nutrient regime that would stimulate biosurfactant production in the oil reservoir
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DEVELOPMENT OF IMPROVED ANAEROBIC GROWTH OF BACILLUS MOJAVENSIS STRAIN JF-2 FOR THE PURPOSE OF IMPROVED ANAEROBIC BIOSURFACTANT PRODUCTION FOR ENHANCED OIL RECOVERY
Our work focuses on the use of microorganisms to recover petroleum hydrocarbons that remain entrapped after current recovery technologies reach their economic limit. Capillary forces between the hydrocarbon and aqueous phases are largely responsible for trapping the hydrocarbons in the pores of the rock and large reductions in the interfacial tension between the hydrocarbon and aqueous phases are needed for hydrocarbon mobilization (1-3, 10, 11). Microorganisms produce a variety of biosurfactants (4), several of which generate the ultra low interfacial tensions needed for hydrocarbon mobilization (4, 5, 8). In particular, the lipopeptide biosurfactant produced by Bacillus mojavensis strain JF-2 reduces the interfacial tension between hydrocarbon and aqueous phases to very low levels (<0.016 mN/m) (8) (9). B. mojavensis JF-2 grows under the environmental conditions found in many oil reservoirs, i. e., anaerobic, NaCl concentrations up to 80 g l{sup -1}, and temperatures up to 45 C (6, 7), making it ideally suited for in situ applications. However, anaerobic growth of B. mojavensis JF-2 was inconsistent and difficult to replicate, which limited its use for in situ applications. Our initial studies revealed that enzymatic digests, such as Proteose Peptone, were required for anaerobic growth of Bacillus mojavensis JF-2. Subsequent purification of the growth-enhancing factor in Proteose Peptone resulted in the identification of the growth-enhancing factor as DNA or deoxyribonucleosides. The addition of salmon sperm DNA, herring sperm DNA, E. coli DNA or synthetic DNA (single or double stranded) to Medium E all supported anaerobic growth of JF-2. Further, we found that JF-2 required all four deoxyribonucleosides (deoxyadeonosine, deoxyguanosine, deoxycytidine and thymidine) for growth under strict anaerobic conditions. The requirement for the deoxyribonucleosides did not occur under aerobic growth conditions. DNA was not used as a sole energy source; sucrose was required for anaerobic growth and biosurfactant production in DNA-supplemented Medium E. In addition to DNA or deoxyribonucleosides, nitrate, amino acids and vitamins were all required for anaerobic growth of JF-2. Bacillus mojavensisT (ABO21191), Bacillus mojavensis, strain ROB2 also required DNA or deoxyribonucleosides for anaerobic growth. The improved anaerobic growth of Bacillus mojavensis JF-2 was a prerequisite for studies that will lead to improved anaerobic biosurfactant production
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Development of Biosurfactant-Mediated Oil Recovery in Model Porous Systems and Computer Simulations of Biosurfactant-Mediated Oil Recovery
Current technology recovers only one-third to one-half of the oil that is originally present in an oil reservoir. Entrapment of petroleum hydrocarbons by capillary forces is a major factor that limits oil recovery (1, 3, 4). Hydrocarbon displacement can occur if interfacial tension (IFT) between the hydrocarbon and aqueous phases is reduced by several orders of magnitude. Microbially-produced biosurfactants may be an economical method to recover residual hydrocarbons since they are effective at low concentrations. Previously, we showed that substantial mobilization of residual hydrocarbon from a model porous system occurs at biosurfactant concentrations made naturally by B. mojavensis strain JF-1 if a polymer and 2,3-butanediol were present (2). In this report, we include data on oil recovery from Berea sandstone experiments along with our previous data from sand pack columns in order to relate biosurfactant concentration to the fraction of oil recovered. We also investigate the effect that the JF-2 biosurfactant has on interfacial tension (IFT). The presence of a co-surfactant, 2,3-butanediol, was shown to improve oil recoveries possibly by changing the optimal salinity concentration of the formulation. The JF-2 biosurfactant lowered IFT by nearly 2 orders of magnitude compared to typical values of 28-29 mN/m. Increasing the salinity increased the IFT with or without 2,3-butanediol present. The lowest interfacial tension observed was 0.1 mN/m. Tertiary oil recovery experiments showed that biosurfactant solutions with concentrations ranging from 10 to 60 mg/l in the presence of 0.1 mM 2,3-butanediol and 1 g/l of partially hydrolyzed polyacrylamide (PHPA) recovered 10-40% of the residual oil present in Berea sandstone cores. When PHPA was used alone, about 10% of the residual oil was recovered. Thus, about 10% of the residual oil recovered in these experiments was due to the increase in viscosity of the displacing fluid. Little or no oil was recovered at biosurfactant concentrations below the critical micelle concentration (about 10 mg/l). Below this concentration, the IFT values were high. At biosurfactant concentrations from 10 to 40 mg/l, the IFT was 1 mN/m. As the biosurfactant concentration increased beyond 40 mg/l, IFT decreased to about 0.1 mN/m. At biosurfactant concentrations in excess of 10 mg/l, residual oil recovery was linearly related to biosurfactant concentration. A modified mathematical model that relates oil recovery to biosurfactant concentration adequately predicted the experimentally observed changes in IFT as a function of biosurfactant concentration
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Development of More Effective Biosurfactants for Enhanced Oil Recovery
The overall goal of this research was to develop effective biosurfactant production for enhanced oil recovery in the United States
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Development of Microorganisms With Improved Transport and Biosurfactant Activity for Enhanced Oil Recovery Progress Report
Diverse microorganisms were screened for biosurfactant production and anaerobic growth at elevated salt concentrations to obtain candidates most suitable for microbial oil recovery. Seventy percent of the 205 strains tested, mostly strains of Bacillus mojavensis, Bacillus subtilis, Bacillus licheniformis, and Bacillus sonorensis, produced biosurfactants aerobically and 41% of the strains had biosurfactant activity greater than Bacillus mojavensis JF-2, the current candidate for oil recovery. Biosurfactant activity varied with the percentage of the 3-hydroxy-tetradecanoate isomers in the fatty acid portion of the biosurfactant. Changing the medium composition by incorporation of different precursors of 3-hydroxy tetradecanoate increased the activity of biosurfactant. The surface tension and critical micelle concentration of 15 different, biosurfactant-producing Bacillus strains was determined individually and in combination with other biosurfactants. Some biosurfactant mixtures were found to have synergistic effect on surface tension (e.g. surface tension was lowered from 41 to 31 mN/m in some cases) while others had a synergistic effect on CMD-1 values. We compared the transport abilities of spores from three Bacillus strains using a model porous system to study spore recovery and transport. Sand-packed columns were used to select for spores or cells with the best transport abilities through brine-saturated sand. Spores of Bacillus mojavensis strains JF-2 and ROB-2 and a natural recombinant, strain C-9, transported through sand at very high efficiencies. The earliest cells/spores that emerged from the column were re-grown, allowed to sporulate, and applied to a second column. This procedure greatly enhanced the transport of strain C-9. Spores with enhanced transport abilities can be easily obtained and that the preparation of inocula for use in MEOR is feasible. Tertiary oil recovery experiments showed that 10 to 40 mg/l of JF-2 biosurfactant in the presence of 0.1 mM 2,3-butanediol and 1 g/l of partially hydrolyzed polyacrylamide (PHPA) recovered 10-40% of residual oil from Berea sandstone cores. When PHPA was used alone, about 10% of the residual oil was recovered. Interfacial tension (IFT) decreased in a stepwise manner as biosurfactant concentration increased with marked reductions in IFT occurring at biosurfactant concentrations of 10 and 40 mg/l. When the biosurfactant concentration was greater than 10 mg/l, residual oil recovery linearly increased with biosurfactant concentration. A mathematical model that relates oil recovery to biosurfactant concentration was modified to include the stepwise changes in IFT as biosurfactant concentrations changes. This model adequately predicted the experimentally observed changes in IFT as a function of biosurfactant concentration. Our work shows that (1) diverse microorganisms produce biosurfactants, (2) nutrient manipulation may provide a mechanism to increase biosurfactant activity, (3) biosurfactant concentrations in excess of the critical micelle concentration recover substantial amounts of residual oil, and (4) equations that describe the effect of the biosurfactant on IFT adequately predict residual oil recovery in sandstone cores
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Development of Microorganisms With Improved Transport and Biosurfactant Activity for Enhanced Oil Recovery Progress Report
Biosurfactants enhance hydrocarbon biodegradation by increasing apparent aqueous solubility or affecting the association of the cell with poorly soluble hydrocarbon. Here, we show that a lipopeptide biosurfactant produced by Bacillus mojavensis strain JF-2 mobilized substantial amounts of residual hydrocarbon from sand-packed columns when a viscosifying agent and a low molecular weight alcohol were present. The amount of residual hydrocarbon mobilized depended on the biosurfactant concentration. One pore volume of cell-free culture fluid with 900 mg/l of the biosurfactant, 10 mM 2,3-butanediol and 1000 mg/l of partially hydrolyzed polyacrylamide polymer mobilized 82% of the residual hydrocarbon. Consistent with the high residual oil recoveries, we found that the bio-surfactant lowered the interfacial tension (IFT) between oil and water by nearly 2 orders of magnitude compared to typical IFT values of 28-29 mN/m. Increasing the salinity increased the IFT with or without 2,3-butanediol present. The lowest interfacial tension observed was 0.1 mN/m. The lipopeptide biosurfactant system may be effective in removing hydrocarbon contamination sources in soils and aquifers and for the recovery of entrapped oil from low production oil reservoirs. Previously, we reported that Proteose peptone was necessary for anaerobic growth and biosurfactant production by B. mojavensis JF-2. The data gathered from crude purification of the growth-enhancing factor in Proteose peptone suggested that it consisted of nucleic acids; however, nucleic acid bases, nucleotides or nucleosides did not replace the requirement for Proteose Peptone. Further studies revealed that salmon sperm DNA, herring sperm DNA, Echerichia coli DNA and synthetic DNA replaced the requirement for Proteose peptone. In addition to DNA, amino acids and nitrate were required for anaerobic growth and vitamins further improved growth. We now have a defined medium that can be used to manipulate growth and biosurfactant production. As an initial step in the search for a better biosurfactant-producing microorganism, 157 bacterial strains were screened for biosurfactant production under both aerobic and anaerobic conditions. A hundred and forty seven strains produced either equal or higher amounts of biosurfactant compared to B. mojavensis JF-2 and the 10 best strains were chosen for further study. In an attempt to increase biosurfactant production, a genetic recombination experiment was conducted by mixing germinating spores of four of the best strains with B. mojavensis JF-2. Biosurfactant production was higher with the mixed spore culture than in the cocultures containing B. mojavensis JF-2 and each of the other 4 strains or in a mixed culture containing all five strains that had not undergone genetic exchange. Four isolates were obtained from the mixed spores culture that gave higher biosurfactant production than any of the original strains. Repetitive sequence-based polymerase chain reaction analysis showed differences in the band pattern for these strains compared to the parent strains, suggesting the occurrence of genetic recombination. We have a large collection of biosurfactant-producing microorganisms and a natural mechanism to improve biosurfactant production in these organisms
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Development of Microorganisms with Improved Transport and Biosurfactant Activity for Enhanced Oil Recovery: Final Report
The project had three objectives: (1) to develop microbial strains with improved biosurfactant properties that use cost-effective nutrients, (2) to obtain biosurfactant strains with improved transport properties through sandstones, and (3) to determine the empirical relationship between surfactant concentration and interfacial tension and whether in situ reactions kinetics and biosurfactant concentration meets appropriate engineering design criteria. Here, we show that a lipopeptide biosurfactant produced by Bacillus mojavensis strain JF-2 mobilized substantial amounts of residual hydrocarbon from sand-packed columns and Berea sandstone cores when a viscosifying agent and a low molecular weight alcohol were present. The amount of residual hydrocarbon mobilized depended on the biosurfactant concentration. Tertiary oil recovery experiments showed that 10 to 40 mg/l of JF-2 biosurfactant in the presence of 0.1 mM 2,3-butanediol and 1 g/l of partially hydrolyzed polyacrylamide (PHPA) recovered 10-40% of residual oil from Berea sandstone cores. Even low biosurfactant concentrations (16 mg/l) mobilized substantial amounts of residual hydrocarbon (29%). The bio-surfactant lowered IFT by nearly 2 orders of magnitude compared to typical IFT values of 28-29 mN/m. Increasing the salinity increased the IFT with or without 2,3-butanediol present. The lowest interfacial tension observed was 0.1 mN/m. A mathematical model that relates oil recovery to biosurfactant concentration was modified to include the stepwise changes in IFT as biosurfactant concentrations changes. This model adequately predicted the experimentally observed changes in IFT as a function of biosurfactant concentration. Theses data show that lipopeptide biosurfactant systems may be effective in removing hydrocarbon contamination sources in soils and aquifers and for the recovery of entrapped oil from low production oil reservoirs. Diverse microorganisms were screened for biosurfactant production and anaerobic growth at elevated salt concentrations to obtain candidates most suitable for microbial oil recovery. Seventy percent of the 205 strains tested, mostly strains of Bacillus mojavensis, Bacillus subtilis, Bacillus licheniformis, and Bacillus sonorensis, produced biosurfactants aerobically and 41% of the strains had biosurfactant activity greater than Bacillus mojavensis JF-2, the current candidate for oil recovery. Biosurfactant activity varied with the percentage of the 3-hydroxy-tetradecanoate isomers in the fatty acid portion of the biosurfactant. Changing the medium composition by incorporation of different precursors of 3-hydroxy tetradecanoate increased the activity of biosurfactant. The surface tension and critical micelle concentration of 15 different, biosurfactant-producing Bacillus strains was determined individually and in combination with other biosurfactants. Some biosurfactant mixtures were found to have synergistic effect on surface tension (e.g. surface tension was lowered from 41 to 31 mN/m in some cases) while others had a synergistic effect on CMD-1 values. We compared the transport abilities of spores from three Bacillus strains using a model porous system to study spore recovery and transport. Sand-packed columns were used to select for spores or cells with the best transport abilities through brine-saturated sand. Spores of Bacillus mojavensis strains JF-2 and ROB-2 and a natural recombinant, strain C-9, transported through sand at very high efficiencies. The earliest cells/spores that emerged from the column were regrown, allowed to sporulate, and applied to a second column. This procedure greatly enhanced the transport of strain C-9. Spores with enhanced transport abilities can be easily obtained and that the preparation of inocula for use in MEOR is feasible. We conducted a push-pull test to study in-situ biosurfactant production by exogenous biosurfactant producers to aid in oil recovery from depleted reservoirs. Five wells from the same formation were used. Two wells received cells and nutrients, two wells were treated with nutrients only, and one well was used as the negative control where only brine was injected. We hypothesized that the wells receiving nutrients and cells treatment would be able to produce biosurfactant in-situ compared to nutrient only treated wells or the negative control. After incubation and a shut-in period to allow in situ growth and metabolism, a series of chemical, microbiological, and molecular analyses were conducted on the produced fluids to obtain evidence for growth, metabolism, and biosurfactant production. Results showed that the wells treated with cells and nutrients indeed produced biosurfactant compared to the other wells as evidenced by the increase in surface activity. Lipopeptide biosurfactants of concentration up to 350 ppm were detected
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Development of More Effective Biosurfactants for Enhanced Oil Recovery
The objectives of this were two fold. First, core displacement studies were done to determine whether microbial processes could recover residual oil at elevated pressures. Second, the importance of biosurfactant production for the recovery of residual oil was studies. In these studies, a biosurfactant-producing, microorganisms called Bacillus licheniformis strain JF-2 was used. This bacterium produces a cyclic peptide biosurfactant that significantly reduces the interfacial tension between oil and brine (7). The use of a mutant deficient in surfactant production and a mathematical MEOR simulator were used to determine the major mechanisms of oil recovery by these two strains