Waste to bioproduct conversion with undefined mixed cultures: the carboxylate platform.

Our societies generate increasing volumes of organic wastes. Considering that we also need alternatives to oil, an opportunity exists to extract liquid fuels or even industrial solvents from these abundant wastes. Anaerobic undefined mixed cultures can handle the complexity and variability of organic wastes, which produces carboxylates that can be efficiently converted to useful bioproducts. However, to date, barriers, such as inefficient liquid product separation and persistence of methanogens, have prevented the production of bioproducts other than methane. Here, we discuss combinations of biological and chemical pathways that comprise the 'carboxylate platform', which is used to convert waste to bioproducts. To develop the carboxylate platform into an important system within biorefineries, we must understand the kinetic and thermodynamic possibilities of anaerobic pathways, understand the ecological principles underlying pathway alternatives, and develop superior separation technologies.

Nutrient and oxygen concentrations within the sediments of an Alaskan beach polluted with the Exxon Valdez oil spill.

Measurements of the background concentrations of nutrients, dissolved oxygen (DO), and salinity were obtained from a beach that has oil from the Exxon Valdez oil spill in 1989. Two transects were set across the beach, one passed through an oil patch while the other transect was clean. Three pits were dug in each transect, and they ranged in depth from 0.9 to 1.5 m. The DO was around 1.0 mg L(-1) at oiled pits and larger than 5 mg L(-1) at clean pits. The average nutrient concentrations in the beach were 0.39 mg-N L(-1) and 0.020 mg-P L(-1). Both concentrations are lower than optimal values for oil biodegradation (2 to 10 mg-N L(-1) and 0.40 to 2.0 mg-P L(-1)), which suggests that they are both limiting factors for biodegradation. The lowest nitrate and DO values were found in the oiled pits, leading to the conclusion that microbial oil consumption was probably occurring under anoxic conditions and was associated to denitrification. We present evidence that the oxygen level may be a major factor limiting oil biodegradation in the beaches.

Microbial reduction of Fe(III) in hematite nanoparticles by Geobacter sulfurreducens.

The rates of microbial Fe(III) reduction of three sizes of hematite nanoparticles by Geobacter sulfurreducens were measured under two H2 partial pressures (0.01 and 1 atm) and three pH (7.0, 7.5, and 8.0) conditions. Hematite particles with mean primary particle sizes of 10, 30, and 50 nm were synthesized by a novel aerosol method that allows tight control of the particle size distribution. The mass-normalized reduction rates of the 10 and 30 nm particles were comparable to each other and higher than the rate for the 50 nm particles. However, the surface area-normalized rate was highest for the 30 nm particles. Consistent with a previously published model, the reduction rates are likely to be proportional to the bacteria-hematite contact area and not to the total hematite surface area. Surface area-normalized iron reduction rates were higher than those reported in previous studies, which may be due to the sequestration of Fe(II) through formation of vivianite. Similar initial reduction rates were observed under all pH and H2 conditions studied.

Assessment of sediment toxicity during anaerobic biodegradation of vegetable oil using Microtox and Hyalella azteca bioassays.

The potential ecological impacts of anaerobic degradation of vegetable oil on freshwater sediments were investigated. Sediment toxicity was evaluated using two regulatory biotests: the Microtox Solid Phase Test and an amphipod (Hyalella azteca) bioassay. The results of the Microtox test showed that the toxicity of the vegetable-oil-contaminated sediments (about 17-33 g oil/kg dry sediments) increased after 2 weeks incubation and then decreased to near background levels after incubation for 8 weeks under anaerobic conditions. The amphipod toxicity bioassay showed that the toxicity of fresh contaminated sediments decreased over time and returned to background levels within 8 weeks. These results suggest that the impact of vegetable oils on organisms within sediments may be limited. To account for the significance of environmental conditions, additional studies over a wide range of incubation conditions (e.g., temperature, nutrient concentration) and other test organisms at various trophic levels are recommended for both acute and chronic toxicity assessment.

Effects of ferric hydroxide on methanogenesis from lipids and long-chain fatty acids in anaerobic digestion.

The addition of ferric hydroxide to sludge from a municipal anaerobic digester stimulated the rate of methanogenesis from canola oil when the initial oil concentration was high (4600 mg/L; P < 0.002), but not when it was low (920 mg/L; P > 0.05). Similar trends were observed when oleic acid, a fatty acid that is a major component of canola oil triglycerides, was provided, but the effects were statistically significant only when the initial concentration of ferric hydroxide was also high (18 g/L; P = 0.015). Iron reduction occurred when ferric hydroxide was added to microcosms containing anaerobic digester sludge, but the extent of ferrous iron production was much less in acetate-amended microcosms than in those that were provided with canola oil or oleic acid. Methanogenesis and acetate consumption were completely inhibited when the initial acetate concentration was approximately 5000 mg/L, regardless of the initial ferric hydroxide concentration. The main effect of ferric hydroxide in this system appears to have been a result of stimulation of the rate of fatty acid oxidation.

Effect of iron on the sensitivity of hydrogen, acetate, and butyrate metabolism to inhibition by long-chain fatty acids in vegetable-oil-enriched freshwater sediments.

Freshwater sediment microbial communities enriched by growth on vegetable oil in the presence of a substoichiometric amount of ferric hydroxide (sufficient to accept about 12% of the vegetable-oil-derived electrons) degrade vegetable oil to methane faster than similar microbial communities that develop when sediments are enriched by growth on vegetable oil in the absence of ferric hydroxide. This study examined the effects of enrichment in the presence of Fe(III) on the fatty-acid sensitivity of several important members of anaerobic triglyceride-degrading microbial communities in freshwater sediments. The fatty-acid sensitivity of three groups of microorganisms-hydrogenotrophic methanogens, acetate consumers, and hydrogen-producing acetogens-were investigated by comparing the rates of hydrogen, acetate, or butyrate consumption in the presence and absence of oleic acid. Methanogenesis from hydrogen was not affected by sediment enrichment conditions or by the presence of oleic acid, suggesting that hydrogenotrophic methanogens were insensitive to fatty acid inhibition in these sediments. Oleic acid inhibited the anaerobic degradation rates of acetate and butyrate by 38% and 63%, respectively, but enrichment in the presence of Fe(III) eliminated the fatty-acid sensitivity of acetate degradation and reduced the sensitivity of butyrate degradation by about half. These results suggest that iron-reducing bacteria may provide an alternative pathway through which vegetable oil can be converted to methane in anaerobic freshwater sediments.

Anaerobic biodegradation of vegetable oil and its metabolic intermediates in oil-enriched freshwater sediments.

Anaerobic biodegradation of vegetable oil in freshwater sediments is strongly inhibited by high concentrations of oil, but the presence of ferric hydroxide relieves the inhibition. The effect of ferric hydroxide is not due to physical or chemical interactions with long-chain fatty acids (LCFAs) that are produced as intermediates during metabolism of vegetable-oil triglycerides. The anaerobic biodegradation of canola oil and mixtures of acetic and oleic acids, two important intermediates of vegetable-oil metabolism, were investigated using sediments enriched on canola oil under methanogenic and iron-reducing conditions to determine whether the effect of ferric hydroxide has a biological basis. Sediments enriched under both conditions rapidly and completely converted canola oil to methane when the initial oil concentration was relatively low (1.9 g oil/kg sediments), but the biotransformation was strongly inhibited in sediments enriched under methanogenic conditions when the initial concentration was 19 g/kg (< 30% of the oil-derived electron equivalents were transferred to methane in a 420-day incubation period). Sediments enriched under iron-reducing conditions, however, completely transformed canola oil to methane in about 250 days at this initial oil concentration. The anaerobic biotransformation of mixtures of acetate and oleic acid followed a similar pattern: the rate and extent of conversion of these electron-donor substrates to methane was always higher in sediments enriched under iron-reducing than under methanogenic conditions. These results suggest that enrichment on canola oil in the presence of ferric hydroxide selects a microbial community that is less sensitive to inhibition by LCFAs than the community that develops during enrichment under methanogenic conditions.

Effects of ferric hydroxide on the anaerobic biodegradation kinetics and toxicity of vegetable oil in freshwater sediments.

Biodegradation of vegetable oil in freshwater sediments exhibits self-inhibitory characteristics when it occurs under methanogenic conditions but not under iron-reducing conditions. The basis of the protective effect of iron was investigated by comparing its effects on oil biodegradation rate and the toxicity of oil-amended sediments to those of clay and calcium, which reduce the toxicity of oil-derived long-chain fatty acids by adsorption and precipitation, respectively. Kinetic parameters for an integrated mixed-second-order model were estimated by nonlinear regression using cumulative methane production as the response variable and used to compare the effects of the three treatment factors on the rate of oil biodegradation. Ferric hydroxide was the only factor that significantly (P<0.05) increased the rate of methane production from canola oil, whereas calcium significantly reduced the oil biodegradation rate. Measurement of sediment toxicity using the Microtox Solid-Phase Test showed that inhibitory products formed within 5 days of oil addition, but the sediment toxicity decreased over time as the extent of oil mineralization increased. None of the other amendments significantly reduced the toxicity of oil-containing sediments. Since ferric hydroxide stimulated the rate of oil biodegradation without affecting the toxicity of oiled sediments, it must operate through a mechanism that is different from those previously described for clay and calcium.

Production of bioenergy and biochemicals from industrial and agricultural wastewater.

The building of a sustainable society will require reduction of dependency on fossil fuels and lowering of the amount of pollution that is generated. Wastewater treatment is an area in which these two goals can be addressed simultaneously. As a result, there has been a paradigm shift recently, from disposing of waste to using it. There are several biological processing strategies that produce bioenergy or biochemicals while treating industrial and agricultural wastewater, including methanogenic anaerobic digestion, biological hydrogen production, microbial fuel cells and fermentation for production of valuable products. However, there are also scientific and technical barriers to the implementation of these strategies.