Abstract

Abstract We are trying to develop a methane-producing system using indigenous microbes in depleted oil fields as a new microbial enhanced oil recovery process. In particular, we aim to combine a microbial conversion of the residual oil into methane with the geological sequestration of carbon dioxide. The mechanism is as follows: Hydrocarbon-degrading bacteria are harnessed to produce hydrogen and/or acetate from residual oil in the depleted oil reservoir. Then, methane-producing microbes (methanogens) utilize the produced acetate or hydrogen and carbon dioxide, which is injected for geological sequestration, to generate methane. We successfully isolated hydrogen- and methane-producing microbes (hydrogen-producing bacteria and methanogens) from oil fields (Yabase and other oil fields) in Japan. Our analysis of microbial cultures incubated under high temperature and high pressure, the condition similar to in situ petroleum reservoir conditions, revealed that indigenous microbes in the reservoir brine are capable of generating methane by utilizing crude oil and carbon dioxide. Consumption/production rate of gases (methane and carbon dioxide) and acetic acid indicated that the methane production under reservoir conditions is likely mediated through two major pathways; the acetoclastic (acetic-acid utilizing) and the hydrogenotrophic (hydrogen and carbon-dioxide utilizing) pathways. Furthermore, by analyzing methane-producing ability of isolated microbes, we found that the syntrophic cooperation between hydrogen-producing bacteria and methanogens was critical for the methane producing under the reservoir condition. 0%.tures with carbon dioxideent Strikingly, addition of carbon dioxide accelerated methane production of the cultures. The methane production rate of the cultures, in which high concentration (10%) of carbon dioxide was supplied into the head spaces, was 0.30 mmol/L/Day. On the other hand, the cultures without the addition of carbon dioxide showed the methane production rate of 0.12 mmol/L/Day, significantly slower (ca. 40%) than the production rate of the cultures with carbon dioxide. These results suggested that addition (injection) of carbon dioxide into reservoirs might accelerate the microbial methane production. We further investigated the methanogenic communities and pathways in petroleum reservoirs by incubating the reservoir brine from the Yabase oil field, combined with radiotracer experiments and molecular biological analyses. The brine samples were incubated without exogenous-nutrient supplementation under the high-temperature and high-pressure condition (the in-reservoir condition). The radiotracer analysis (using 14C-biocarbonate and 14C-acetate) indicated that the methane production rate of hydrogenotrophic methanogenesis was 50-fold higher than that of acetoclastic methanogenesis, suggesting dominance of methane production by syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis in reservoir. In this study, we assessed the rate of oil biodegradation coupled with methanogenesis by using 14C-labeled toluene and hexadecane as tracers. The analysis revealed that the rate was very low, being only about one thousandth of that of the hydrogenotrophic methanogenesis. We are currently trying to enhance the crude-oil biodegradation for effective conversion of crude oil to methane. Our goal is to establish effective microbial conversion system from residual oil into methane in depleted oil fields as a new EOR technology.

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