Background: Large-scale algal biofuel production has been limited, among other factors, by the av... more Background: Large-scale algal biofuel production has been limited, among other factors, by the availability of inorganic carbon in the culture medium at concentrations higher than achievable with atmospheric CO 2. Life cycle analyses have concluded that costs associated with supplying CO 2 to algal cultures are significant contributors to the overall energy consumption. Results: A two-phase optimal growth and lipid accumulation scenario is presented, which (1) enhances the growth rate and (2) the triacylglyceride (TAG) accumulation rate in the oleaginous Chlorophyte Chlorella vulgaris strain UTEX 395, by growing the organism in the presence of low concentrations of NaHCO 3 (5 mM) and controlling the pH of the system with a periodic gas sparge of 5 % CO 2 (v/v). Once cultures reached the desired cell densities, which can be "fine-tuned" based on initial nutrient concentrations, cultures were switched to a lipid accumulation metabolism through the addition of 50 mM NaHCO 3. This two-phase approach increased the specific growth rate of C. vulgaris by 69 % compared to cultures sparged continuously with 5 % CO 2 (v/v); further, biomass productivity (g L −1 day −1) was increased by 27 %. Total biodiesel potential [assessed as total fatty acid methyl ester (FAME) produced] was increased from 53.3 to 61 % (FAME biomass −1) under the optimized conditions; biodiesel productivity (g FAME L −1 day −1) was increased by 7.7 %. A bicarbonate salt screen revealed that American Chemical Society (ACS) and industrial grade NaHCO 3 induced the highest TAG accumulation (% w/w), whereas Na 2 CO 3 did not induce significant TAG accumulation. NH 4 HCO 3 had a negative effect on cell health presumably due to ammonia toxicity. The raw, unrefined form of trona, NaHCO 3 •Na 2 CO 3 (sodium sesquicarbonate) induced TAG accumulation, albeit to a slightly lower extent than the more refined forms of sodium bicarbonate. Conclusions: The strategic addition of sodium bicarbonate was found to enhance growth and lipid accumulation rates in cultures of C. vulgaris, when compared to traditional culturing strategies, which rely on continuously sparging algal cultures with elevated concentrations of CO 2(g). This work presents a two-phased, improved photoautotrophic growth and lipid accumulation approach, which may result in an overall increase in algal biofuel productivity.
Research on microbially induced calcite precipitation (MICP) is reported. MICP may serve to reduc... more Research on microbially induced calcite precipitation (MICP) is reported. MICP may serve to reduce near-wellbore permeability, reduce CO 2related corrosion, and lower the risk of unwanted migration of CO 2 or other fluids. MICP research on the lab scale has demonstrated the ability to seal sandstone cores using injection strategies engineered to control precipitation. Experimentation was also aimed at transitioning MICP strategies for field implementation. MICP was evaluated in the field in a hydraulically fractured sandstone formation at a Walker County, Alabama well. The field experiment resulted in greatly reduced injectivity indicating that the fractured formation was plugged after MICP treatment.
Various U. S. Department of Energy (DOE) low and medium-level radioactive waste sites contain mix... more Various U. S. Department of Energy (DOE) low and medium-level radioactive waste sites contain mixtures of heavy metals, radionuclides and assorted organic materials. Over time, water infiltrates the wastes, and releases metals and radionuclides causing transport into the surrounding environment. We propose that fermentative microorganisms are active in these sites and may control metal and radionuclide migration from source zones (Figure 1). The following overarching hypothesis will drive our research: 'Metals and radionuclides can be mobilized by infiltration of water into waste storage sites. Microbial communities of lignocellulose degrading and fermenting microorganisms present in the subsurface of contaminated DOE sites can significantly impact migration by directly reducing and immobilizing metals and radionuclides while degrading complex organic matter to low molecular weight organic compounds. These low molecular weight organic compounds can increase metal and radionuclide mobility by chelation (i.e., certain organic acids) or decrease mobility by stimulating respiratory metal reducing microorganisms.' The objective of our research is to determine the effect of carbon and energy flow through simulated waste environments on metal and radionuclide migration from waste pits and trenches across the DOE complex. Metals and radionuclides can be mobilized by infiltration of water into waste storage sites. Cellulolytic and non-cellulolytic fermentative microorganisms have been chosen as the focus of this research because their activity is a critical first step that we hypothesize will control subsequent fate and transport in contaminated natural systems. Microbial communities of lignocellulose degrading and fermenting microorganisms present in the subsurface of contaminated DOE sites can significantly impact migration by directly reducing and immobilizing metals and radionuclides while degrading complex organic matter to low molecular weight organic compounds. These low molecular weight organic acids and alcohols can increase metal and radionuclide mobility by chelation (i.e., certain organic acids) or decrease mobility by stimulating respiratory metal reducing microorganisms.
The collaborative project was designed to evaluate the possibility developing a subsurface remedi... more The collaborative project was designed to evaluate the possibility developing a subsurface remediation technology for mixed wastes at Department of Energy sites using a group of common soil bacteria of the genus Cellulomonas. We have been gaining a better understanding of microbial transformation of chromium, uranium, iron minerals, and trinitrotoluene (TNT) by Cellulomonas spp. in simulated subsurface environments.
The Heart Lake Geyser Basin (HLGB) is remotely located at the base of Mount Sheridan in southern ... more The Heart Lake Geyser Basin (HLGB) is remotely located at the base of Mount Sheridan in southern Yellowstone National Park (YNP), Wyoming, USA and is situated along Witch Creek and the northwestern shore of Heart Lake. Likely because of its location, little is known about the microbial community structure of springs in the HLGB. Bacterial and archaeal populations were monitored via small subunit (SSU) rRNA gene pyrosequencing over 3 years in 3 alkaline (pH 8.5) hot springs with varying temperatures (44°C, 63°C, 75°C). The bacterial populations were generally stable over time, but varied by temperature. The dominant bacterial community changed from moderately thermophilic and photosynthetic members (Cyanobacteria and Chloroflexi) at 44°C to a mixed photosynthetic and thermophilic community (Deinococcus-Thermus) at 63°C and a non-photosynthetic thermophilic community at 75°C. The archaeal community was more variable across time and was predominantly a methanogenic community in the 44 ...
ABSTRACT Inorganic carbon is a fundamental component for microalgal lipid biosynthesis. Understan... more ABSTRACT Inorganic carbon is a fundamental component for microalgal lipid biosynthesis. Understanding how the concentration and speciation of dissolved inorganic carbon (DIC) influences lipid metabolism in microalgae may help researchers optimize the production of these high value metabolites. Using relatively straight forward methods for quantifying free fatty acids (FFAs), mono- (MAG), di- (DAG), tri-acylglycerides (TAG), and total cellular fatty acids (FAME), lipid profiles over time were established for Chlamydomonas reinhardtii when grown under three unique inorganic carbon regimes. Specifically, cultures sparged with atmospheric air were compared to cultures which were sparged with 5% CO2 (v/v) and cultures supplemented with 50 mM NaHCO3 just prior to medium nitrogen depletion. All three conditions exhibited similar lipid profiles prior to nitrogen depletion in the medium, with FFA and MAG being the predominant lipid metabolites. However, these precursors were quickly reallocated into DAG and subsequently TAG after nitrogen depletion. C16 DAG did not accumulate significantly in any of the treatments, whereas the C18 DAG content increased throughout both exponential growth and stationary phase. C16 and C18 TAG began to accumulate after nitrogen depletion, with C16 TAG contributing the most to overall TAG content. C16 fatty acids exhibited a shift towards saturated C16 fatty acids after nitrogen depletion. Results provide insight into inorganic carbon partitioning into lipid compounds and how the organism's lipid metabolism changes due to N-deplete culturing and inorganic carbon source availability. The methodologies and findings presented here may be adapted to other organisms with high industrial relevance.
Microalgal biofilms grown to evaluate potential nutrient removal options for wastewaters and feed... more Microalgal biofilms grown to evaluate potential nutrient removal options for wastewaters and feedstock for biofuels production were studied to determine the influence of bicarbonate amendment on their growth, nutrient uptake capacity, and lipid accumulation after nitrogen starvation. No significant differences in growth rates, nutrient removal, or lipid accumulation were observed in the algal biofilms with or without bicarbonate amendment. The biofilms possibly did not experience carbon-limited conditions because of the large reservoir of dissolved inorganic carbon in the medium. However, an increase in photosynthetic rates was observed in algal biofilms amended with bicarbonate. The influence of bicarbonate on photosynthetic and respiration rates was especially noticeable in biofilms that experienced nitrogen stress. Medium nitrogen depletion was not a suitable stimulant for lipid production in the algal biofilms and as such, focus should be directed toward optimizing growth and biomass productivities to compensate for the low lipid yields and increase nutrient uptake.
Knowledge of taxis (directed swimming) in the Archaea is currently expanding through identificati... more Knowledge of taxis (directed swimming) in the Archaea is currently expanding through identification of novel receptors, effectors, and proteins involved in signal transduction to the flagellar motor. Although the ability for biological cells to sense and swim toward hydrogen gas has been hypothesized for many years, this capacity has yet to be observed and demonstrated. Here we show that the average swimming velocity increases in the direction of a source of hydrogen gas for the methanogen, Methanococcus maripaludis using a capillary assay with anoxic gas-phase control and time-lapse microscopy. The results indicate that a methanogen couples motility to hydrogen concentration sensing and is the first direct observation of hydrogenotaxis in any domain of life. Hydrogenotaxis represents a strategy that would impart a competitive advantage to motile microorganisms that compete for hydrogen gas and would impact the C, S and N cycles.
ABSTRACT If CO2 is injected in deep geological formations it is important that the receiving form... more ABSTRACT If CO2 is injected in deep geological formations it is important that the receiving formation has sufficient porosity and permeability for storage and transmission and be overlain by a suitable low-permeability cap rock formation. When the resulting CO2 plume encounters a well bore, leakage may occur through various pathways in the “disturbed zone” surrounding the well casing. Gasda et al. , propose a method for determining effective well bore permeability from a field pressure test. If permeability results from such tests prove unacceptably large, strategies for in situ mitigation of potential leakage pathways become important. To be effective, leakage mitigation methods must block leakage pathways on timescales longer than the plume will be mobile, be able to be delivered without causing well screen plugging, and be resistant to supercritical CO2 (ScCO2) challenges. Traditional mitigation uses cement, a viscous fluid that requires a large enough aperture for delivery and that also must bond to the surrounding surfaces in order to be effective. Technologies that can be delivered via low viscosity fluids and that can effectively plug small aperture pathways, or even the porous rock surrounding the well could have significant advantages for some leakage scenarios.
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
In order to develop subsurface CO2 storage as a viable engineered mechanism to reduce concentrati... more In order to develop subsurface CO2 storage as a viable engineered mechanism to reduce concentrations of atmospheric CO2, any potential ¡°leakage¡± of injected supercritical CO2 (scCO2) from the ground to the atmosphere must be reduced. Here, we investigate the utility of biofilms, which are microorganism assemblages firmly attached to a surface, as a means of reducing scCO2 leakage. Firstly, experiments
There are four trapping mechanisms proposed to play significant roles in the deep geologic seques... more There are four trapping mechanisms proposed to play significant roles in the deep geologic sequestration of CO2: i) formation trapping, ii) capillary trapping, iii) solubility trapping, and iv) mineral trapping. Our research has shown that microbial biofilms are capable of enhancing formation trapping, solubility trapping, and mineral trapping under conditions found in brine aquifers targeted for geologic carbon sequestration. We
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
Background: Large-scale algal biofuel production has been limited, among other factors, by the av... more Background: Large-scale algal biofuel production has been limited, among other factors, by the availability of inorganic carbon in the culture medium at concentrations higher than achievable with atmospheric CO 2. Life cycle analyses have concluded that costs associated with supplying CO 2 to algal cultures are significant contributors to the overall energy consumption. Results: A two-phase optimal growth and lipid accumulation scenario is presented, which (1) enhances the growth rate and (2) the triacylglyceride (TAG) accumulation rate in the oleaginous Chlorophyte Chlorella vulgaris strain UTEX 395, by growing the organism in the presence of low concentrations of NaHCO 3 (5 mM) and controlling the pH of the system with a periodic gas sparge of 5 % CO 2 (v/v). Once cultures reached the desired cell densities, which can be "fine-tuned" based on initial nutrient concentrations, cultures were switched to a lipid accumulation metabolism through the addition of 50 mM NaHCO 3. This two-phase approach increased the specific growth rate of C. vulgaris by 69 % compared to cultures sparged continuously with 5 % CO 2 (v/v); further, biomass productivity (g L −1 day −1) was increased by 27 %. Total biodiesel potential [assessed as total fatty acid methyl ester (FAME) produced] was increased from 53.3 to 61 % (FAME biomass −1) under the optimized conditions; biodiesel productivity (g FAME L −1 day −1) was increased by 7.7 %. A bicarbonate salt screen revealed that American Chemical Society (ACS) and industrial grade NaHCO 3 induced the highest TAG accumulation (% w/w), whereas Na 2 CO 3 did not induce significant TAG accumulation. NH 4 HCO 3 had a negative effect on cell health presumably due to ammonia toxicity. The raw, unrefined form of trona, NaHCO 3 •Na 2 CO 3 (sodium sesquicarbonate) induced TAG accumulation, albeit to a slightly lower extent than the more refined forms of sodium bicarbonate. Conclusions: The strategic addition of sodium bicarbonate was found to enhance growth and lipid accumulation rates in cultures of C. vulgaris, when compared to traditional culturing strategies, which rely on continuously sparging algal cultures with elevated concentrations of CO 2(g). This work presents a two-phased, improved photoautotrophic growth and lipid accumulation approach, which may result in an overall increase in algal biofuel productivity.
Research on microbially induced calcite precipitation (MICP) is reported. MICP may serve to reduc... more Research on microbially induced calcite precipitation (MICP) is reported. MICP may serve to reduce near-wellbore permeability, reduce CO 2related corrosion, and lower the risk of unwanted migration of CO 2 or other fluids. MICP research on the lab scale has demonstrated the ability to seal sandstone cores using injection strategies engineered to control precipitation. Experimentation was also aimed at transitioning MICP strategies for field implementation. MICP was evaluated in the field in a hydraulically fractured sandstone formation at a Walker County, Alabama well. The field experiment resulted in greatly reduced injectivity indicating that the fractured formation was plugged after MICP treatment.
Various U. S. Department of Energy (DOE) low and medium-level radioactive waste sites contain mix... more Various U. S. Department of Energy (DOE) low and medium-level radioactive waste sites contain mixtures of heavy metals, radionuclides and assorted organic materials. Over time, water infiltrates the wastes, and releases metals and radionuclides causing transport into the surrounding environment. We propose that fermentative microorganisms are active in these sites and may control metal and radionuclide migration from source zones (Figure 1). The following overarching hypothesis will drive our research: 'Metals and radionuclides can be mobilized by infiltration of water into waste storage sites. Microbial communities of lignocellulose degrading and fermenting microorganisms present in the subsurface of contaminated DOE sites can significantly impact migration by directly reducing and immobilizing metals and radionuclides while degrading complex organic matter to low molecular weight organic compounds. These low molecular weight organic compounds can increase metal and radionuclide mobility by chelation (i.e., certain organic acids) or decrease mobility by stimulating respiratory metal reducing microorganisms.' The objective of our research is to determine the effect of carbon and energy flow through simulated waste environments on metal and radionuclide migration from waste pits and trenches across the DOE complex. Metals and radionuclides can be mobilized by infiltration of water into waste storage sites. Cellulolytic and non-cellulolytic fermentative microorganisms have been chosen as the focus of this research because their activity is a critical first step that we hypothesize will control subsequent fate and transport in contaminated natural systems. Microbial communities of lignocellulose degrading and fermenting microorganisms present in the subsurface of contaminated DOE sites can significantly impact migration by directly reducing and immobilizing metals and radionuclides while degrading complex organic matter to low molecular weight organic compounds. These low molecular weight organic acids and alcohols can increase metal and radionuclide mobility by chelation (i.e., certain organic acids) or decrease mobility by stimulating respiratory metal reducing microorganisms.
The collaborative project was designed to evaluate the possibility developing a subsurface remedi... more The collaborative project was designed to evaluate the possibility developing a subsurface remediation technology for mixed wastes at Department of Energy sites using a group of common soil bacteria of the genus Cellulomonas. We have been gaining a better understanding of microbial transformation of chromium, uranium, iron minerals, and trinitrotoluene (TNT) by Cellulomonas spp. in simulated subsurface environments.
The Heart Lake Geyser Basin (HLGB) is remotely located at the base of Mount Sheridan in southern ... more The Heart Lake Geyser Basin (HLGB) is remotely located at the base of Mount Sheridan in southern Yellowstone National Park (YNP), Wyoming, USA and is situated along Witch Creek and the northwestern shore of Heart Lake. Likely because of its location, little is known about the microbial community structure of springs in the HLGB. Bacterial and archaeal populations were monitored via small subunit (SSU) rRNA gene pyrosequencing over 3 years in 3 alkaline (pH 8.5) hot springs with varying temperatures (44°C, 63°C, 75°C). The bacterial populations were generally stable over time, but varied by temperature. The dominant bacterial community changed from moderately thermophilic and photosynthetic members (Cyanobacteria and Chloroflexi) at 44°C to a mixed photosynthetic and thermophilic community (Deinococcus-Thermus) at 63°C and a non-photosynthetic thermophilic community at 75°C. The archaeal community was more variable across time and was predominantly a methanogenic community in the 44 ...
ABSTRACT Inorganic carbon is a fundamental component for microalgal lipid biosynthesis. Understan... more ABSTRACT Inorganic carbon is a fundamental component for microalgal lipid biosynthesis. Understanding how the concentration and speciation of dissolved inorganic carbon (DIC) influences lipid metabolism in microalgae may help researchers optimize the production of these high value metabolites. Using relatively straight forward methods for quantifying free fatty acids (FFAs), mono- (MAG), di- (DAG), tri-acylglycerides (TAG), and total cellular fatty acids (FAME), lipid profiles over time were established for Chlamydomonas reinhardtii when grown under three unique inorganic carbon regimes. Specifically, cultures sparged with atmospheric air were compared to cultures which were sparged with 5% CO2 (v/v) and cultures supplemented with 50 mM NaHCO3 just prior to medium nitrogen depletion. All three conditions exhibited similar lipid profiles prior to nitrogen depletion in the medium, with FFA and MAG being the predominant lipid metabolites. However, these precursors were quickly reallocated into DAG and subsequently TAG after nitrogen depletion. C16 DAG did not accumulate significantly in any of the treatments, whereas the C18 DAG content increased throughout both exponential growth and stationary phase. C16 and C18 TAG began to accumulate after nitrogen depletion, with C16 TAG contributing the most to overall TAG content. C16 fatty acids exhibited a shift towards saturated C16 fatty acids after nitrogen depletion. Results provide insight into inorganic carbon partitioning into lipid compounds and how the organism's lipid metabolism changes due to N-deplete culturing and inorganic carbon source availability. The methodologies and findings presented here may be adapted to other organisms with high industrial relevance.
Microalgal biofilms grown to evaluate potential nutrient removal options for wastewaters and feed... more Microalgal biofilms grown to evaluate potential nutrient removal options for wastewaters and feedstock for biofuels production were studied to determine the influence of bicarbonate amendment on their growth, nutrient uptake capacity, and lipid accumulation after nitrogen starvation. No significant differences in growth rates, nutrient removal, or lipid accumulation were observed in the algal biofilms with or without bicarbonate amendment. The biofilms possibly did not experience carbon-limited conditions because of the large reservoir of dissolved inorganic carbon in the medium. However, an increase in photosynthetic rates was observed in algal biofilms amended with bicarbonate. The influence of bicarbonate on photosynthetic and respiration rates was especially noticeable in biofilms that experienced nitrogen stress. Medium nitrogen depletion was not a suitable stimulant for lipid production in the algal biofilms and as such, focus should be directed toward optimizing growth and biomass productivities to compensate for the low lipid yields and increase nutrient uptake.
Knowledge of taxis (directed swimming) in the Archaea is currently expanding through identificati... more Knowledge of taxis (directed swimming) in the Archaea is currently expanding through identification of novel receptors, effectors, and proteins involved in signal transduction to the flagellar motor. Although the ability for biological cells to sense and swim toward hydrogen gas has been hypothesized for many years, this capacity has yet to be observed and demonstrated. Here we show that the average swimming velocity increases in the direction of a source of hydrogen gas for the methanogen, Methanococcus maripaludis using a capillary assay with anoxic gas-phase control and time-lapse microscopy. The results indicate that a methanogen couples motility to hydrogen concentration sensing and is the first direct observation of hydrogenotaxis in any domain of life. Hydrogenotaxis represents a strategy that would impart a competitive advantage to motile microorganisms that compete for hydrogen gas and would impact the C, S and N cycles.
ABSTRACT If CO2 is injected in deep geological formations it is important that the receiving form... more ABSTRACT If CO2 is injected in deep geological formations it is important that the receiving formation has sufficient porosity and permeability for storage and transmission and be overlain by a suitable low-permeability cap rock formation. When the resulting CO2 plume encounters a well bore, leakage may occur through various pathways in the “disturbed zone” surrounding the well casing. Gasda et al. , propose a method for determining effective well bore permeability from a field pressure test. If permeability results from such tests prove unacceptably large, strategies for in situ mitigation of potential leakage pathways become important. To be effective, leakage mitigation methods must block leakage pathways on timescales longer than the plume will be mobile, be able to be delivered without causing well screen plugging, and be resistant to supercritical CO2 (ScCO2) challenges. Traditional mitigation uses cement, a viscous fluid that requires a large enough aperture for delivery and that also must bond to the surrounding surfaces in order to be effective. Technologies that can be delivered via low viscosity fluids and that can effectively plug small aperture pathways, or even the porous rock surrounding the well could have significant advantages for some leakage scenarios.
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
In order to develop subsurface CO2 storage as a viable engineered mechanism to reduce concentrati... more In order to develop subsurface CO2 storage as a viable engineered mechanism to reduce concentrations of atmospheric CO2, any potential ¡°leakage¡± of injected supercritical CO2 (scCO2) from the ground to the atmosphere must be reduced. Here, we investigate the utility of biofilms, which are microorganism assemblages firmly attached to a surface, as a means of reducing scCO2 leakage. Firstly, experiments
There are four trapping mechanisms proposed to play significant roles in the deep geologic seques... more There are four trapping mechanisms proposed to play significant roles in the deep geologic sequestration of CO2: i) formation trapping, ii) capillary trapping, iii) solubility trapping, and iv) mineral trapping. Our research has shown that microbial biofilms are capable of enhancing formation trapping, solubility trapping, and mineral trapping under conditions found in brine aquifers targeted for geologic carbon sequestration. We
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trappi... more The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.
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Papers by Robin Gerlach