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In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.

ABSTRACT: Biosurfactant-mediated oil recovery may be an economic approach for recovery of significant amounts of oil entrapped in reservoirs, but evidence that biosurfactants can be produced in situ at concentrations needed to mobilize oil is lacking. We tested whether two Bacillus strains that produce lipopeptide biosurfactants can metabolize and produce their biosurfactants in an oil reservoir. Five wells that produce from the same Viola limestone formation were used. Two wells received an inoculum (a mixture of Bacillus strain RS-1 and Bacillus subtilis subsp. spizizenii NRRL B-23049) and nutrients (glucose, sodium nitrate, and trace metals), two wells received just nutrients, and one well received only formation water. Results showed in situ metabolism and biosurfactant production. The average concentration of lipopeptide biosurfactant in the produced fluids of the inoculated wells was about 90 mg/liter. This concentration is approximately nine times the minimum concentration required to mobilize entrapped oil from sandstone cores. Carbon dioxide, acetate, lactate, ethanol, and 2,3-butanediol were detected in the produced fluids of the inoculated wells. Only CO(2) and ethanol were detected in the produced fluids of the nutrient-only-treated wells. Microbiological and molecular data showed that the microorganisms injected into the formation were retrieved in the produced fluids of the inoculated wells. We provide essential data for modeling microbial oil recovery processes in situ, including growth rates (0.06 +/- 0.01 h(-1)), carbon balances (107% +/- 34%), biosurfactant production rates (0.02 +/- 0.001 h(-1)), and biosurfactant yields (0.015 +/- 0.001 mol biosurfactant/mol glucose). The data demonstrate the technical feasibility of microbial processes for oil recovery.

SUBMITTER: Youssef N

PROVIDER: S-EPMC1828672 | biostudies-literature | 2007 Feb

REPOSITORIES: biostudies-literature

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In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.

Youssef N N Simpson D R DR Duncan K E KE McInerney M J MJ Folmsbee M M Fincher T T Knapp R M RM

Applied and environmental microbiology 20061215 4

Biosurfactant-mediated oil recovery may be an economic approach for recovery of significant amounts of oil entrapped in reservoirs, but evidence that biosurfactants can be produced in situ at concentrations needed to mobilize oil is lacking. We tested whether two Bacillus strains that produce lipopeptide biosurfactants can metabolize and produce their biosurfactants in an oil reservoir. Five wells that produce from the same Viola limestone formation were used. Two wells received an inoculum (a m ...[more]

PMID: 17172458

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Project description:Subsurface petroleum reservoirs are an important component of the deep biosphere where indigenous microorganisms live under extreme conditions and in isolation from the Earth's surface for millions of years. However, unlike the bulk of the deep biosphere, the petroleum reservoir deep biosphere is subject to extreme anthropogenic perturbation, with the introduction of new electron acceptors, donors and exogenous microbes during oil exploration and production. Despite the fundamental and practical significance of this perturbation, there has never been a systematic evaluation of the ecological changes that occur over the production lifetime of an active offshore petroleum production system. Analysis of the entire Halfdan oil field in the North Sea (32 producing wells in production for 1-15 years) using quantitative PCR, multigenic sequencing, comparative metagenomic and genomic bins reconstruction revealed systematic shifts in microbial community composition and metabolic potential, as well as changing ecological strategies in response to anthropogenic perturbation of the oil field ecosystem, related to length of time in production. The microbial communities were initially dominated by slow growing anaerobes such as members of the Thermotogales and Clostridiales adapted to living on hydrocarbons and complex refractory organic matter. However, as seawater and nitrate injection (used for secondary oil production) delivered oxidants, the microbial community composition progressively changed to fast growing opportunists such as members of the Deferribacteres, Delta-, Epsilon- and Gammaproteobacteria, with energetically more favorable metabolism (for example, nitrate reduction, H2S, sulfide and sulfur oxidation). This perturbation has profound consequences for understanding the microbial ecology of the system and is of considerable practical importance as it promotes detrimental processes such as reservoir souring and metal corrosion. These findings provide a new conceptual framework for understanding the petroleum reservoir biosphere and have consequences for developing strategies to manage microbiological problems in the oil industry.

Project description:Petroleum is, up to this date, an inimitable nonrenewable energy resource. Petroleum leakage, which arises during transport, storage, and refining, is the most important contaminant in the environment, as it produces harm to the surrounding ecosystem. Bioremediation is an efficient method used to treat petroleum hydrocarbon-contaminated soil using indigenous microorganisms. The degradation characteristics for a variety of hydrocarbons (hexane, benzene, gasoline, and diesel) were qualitatively and quantitatively investigated using Bacillus isolates. Microbiological and biochemical methods have been used including isolation of oil-degrading bacteria, enzymatic activities, the determination of physicochemical parameters, biosurfactant production and extraction assay, oil displacement assay, antimicrobial assay of the biosurfactants, and bioremediation kinetics. Consequently, of the 60 isolates capable of degrading different hydrocarbons at fast rates, 34 were suspected to be Bacillus isolates capable of growing in 24 h or 48 h on BH medium supplemented with 2% of hexane, benzene, gasoline, diesel, and olive oil, respectively. Among the 34 isolates, 61% (21/34) are capable of producing biosurfactant-like molecules by using gasoline, 70% (24/34) with diesel oil, 85% (29/34) with hexane, and 82% (28/34) with benzene. It was found that biosurfactant-producing isolates are extractable with HCl (100%), ammonium sulphate (95%), chloroform (95%), and ethanol (100%). Biosurfactants showed stability at 20°C, 37°C, 40°C, and 60°C. Biosurfactant secreted by Bacillus strains has shown an antagonistic effect in Escherichia coli, Shigella flexneri 5a M90T, and Bacillus cereus. The selected isolates could therefore be safely used for biodegradation. Substrate biodegradation patterns by individual isolates were found to significantly differ. The study shows that benzene was degraded faster, followed by hexane, gasoline, and finally diesel. The Bacillus consortium used can decrease hydrocarbon content from 195 to 112 (g/kg) in 15 days.

Dataset Information

In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.

Publications

In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.

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