Kirsten J. J. Steinbusch

Bioelectrochemical Ethanol Production through Mediated Acetate Reduction by Mixed Cultures

Biological acetate reduction with hydrogen is a potential method to convert wet biomass waste into ethanol. Since the ethanol concentration and reaction rates are low, this research studies the feasibility of using an electrode, in stead of hydrogen, as an electron donor for biological acetate reduction in conjunction of an electron mediator. Initially, the effect of three selected mediators on metabolic flows during acetate reduction with hydrogen was explored; subsequently, the best performing mediator was used in a bioelectrochemical system to stimulate acetate reduction at the cathode with mixed cultures at an applied cathode potential of -550 mV. In the batch test, methyl viologen (MV) was found to accelerate ethanol production 6-fold and increased ethanol concentration 2-fold to 13.5 +/- 0.7 mM compared to the control. Additionally, MV inhibited n-butyrate and methane formation, resulting in high ethanol production efficiency (74.6 +/- 6%). In the bioelectrochemical system, MV addition to an inoculated cathode led directly to ethanol production (1.82 mM). Hydrogen was coproduced at the cathode (0.0035 Nm(3) hydrogen m(-2) d(-1)), so it remained unclear whether acetate was reduced to ethanol by electrons supplied by the mediator or by hydrogen. As MV reacted irreversibly at the cathode, ethanol production stopped after 5 days.

Biological formation of caproate and caprylate from acetate: fuel and chemical production from low grade biomass

This research introduces an alternative mixed culture fermentation technology for anaerobic digestion to recover valuable products from low grade biomass. In this mixed culture fermentation, organic waste streams are converted to caproate and caprylate as precursors for biodiesel or chemicals. It was found that acetate, as the main intermediate of anaerobic digestion, can be elongated to medium chain fatty acids with six and eight carbon atoms. Mixed microbial communities were able to produce 8.17 g l−1 caproate and 0.32 g l−1 caprylate under methanogenesis-suppressed conditions in a stable batch reactor run. The highest production rate was 25.6 mM C caproate per day with a product yield of 0.6 mol C per mol C. This elongation process occurred with both ethanol and hydrogen as electron donors, demonstrating the flexibility of the process. Microbial characterization revealed that the microbial populations were stable and dominated by relatives of Clostridium kluyveri.

Alcohol production through volatile fatty acids reduction with hydrogen as electron donor by mixed cultures.

In this research we demonstrated a new method to produce alcohols. It was experimentally feasible to produce ethanol, propanol and butanol from solely volatile fatty acids (VFAs) with hydrogen as electron donor. In batch tests, VFAs such as acetic, propionic and butyric acids were reduced by mixed microbial cultures with a headspace of 1.5 bar of hydrogen. Observed alcohol concentrations were 3.69+/-0.25 mM of ethanol, 8.08+/-0.85 mM of propanol and 3.66+/-0.05 mM of n-butanol. The conversion efficiency based on the electron balance was 55.1+/-5.6% with acetate as substrate, 50.3+/-4.7% with propionate and 46.7+/-2.2% with n-butyrate. Methane was the most predominant by-product in each batch experiment, 33.6+/-9.6% of VFA and hydrogen was converted to methane with acetate as substrate; which was 27.1+/-7.1% with propionate and 36.6+/-2.2% with n-butyrate. This VFAs reducing renewable fuel production process does not require carbohydrates like fermentable sugars, but uses biomass with high water content or low sugar content that is unsuitable as feedstock for current fermentation processes. This so-called low-grade biomass is abundantly present in many agricultural areas and is economically very attractive feedstock for the production of biofuels.

Chain Elongation with Reactor Microbiomes: Open-Culture Biotechnology To Produce Biochemicals

Chain elongation into medium-chain carboxylates, such as n-caproate and n-caprylate, with ethanol as an electron donor and with open cultures of microbial consortia (i.e., reactor microbiomes) under anaerobic conditions is being developed as a biotechnological production platform. The goal is to use the high thermodynamic efficiency of anaerobic fermentation to convert organic biomass or organic wastes into valuable biochemicals that can be extracted. Several liter-scale studies have been completed and a first pilot-plant study is underway. However, the underlying microbial pathways are not always well understood. In addition, an interdisciplinary approach with knowledge from fields ranging from microbiology and chemical separations to biochemistry and environmental engineering is required. To bring together research from different fields, we reviewed the literature starting with the microbiology and ending with the bioprocess engineering studies that already have been performed. Because understanding the microbial pathways is so important to predict and steer performance, we delved into a stoichiometric and thermodynamic model that sheds light on the effect of substrate ratios and environmental conditions on product formation. Finally, we ended with an outlook.

Chain elongation of acetate and ethanol in an upflow anaerobic filter for high rate MCFA production

Recently, interest has regained for medium chain fatty acids (MCFAs) as a low cost feedstock for bio-based chemical and fuel production processes. To become cost-effective, the volumetric MCFA production rate by chain elongation should increase to comparable rates of other fermentation processes. We investigate the MCFA production process at a hydraulic retention time of 17 h in an upflow anaerobic filter to improve the volumetric MCFA production rate. This approach resulted in a MCFA production with a volumetric production rate of 16.6 g l(-1) d(-1), which is more than seven times higher than the current production rate. Moreover the rate is now in the range of other fermentation processes like methane, butanol and ethanol production. Increasing the ethanol load lead to higher volumetric production rates and a high MCFA selectivity of 91%. During operation, methane percentages lower than 0.1% were detected in the headspace of reactor.

Improving medium chain fatty acid productivity using chain elongation by reducing the hydraulic retention time in an upflow anaerobic filter

The objective of this investigation was to further increase the medium chain fatty acid (MCFA) production rate by reducing the hydraulic retention time (HRT) in an upflow anaerobic filter. The results showed that the volumetric MCFA production rate was increased to 57.4 g MCFA l(-1) d(-1), more than 3 times higher than previous work. Despite the lower MCFA concentrations at 4h HRT, the MCFA selectivity remained above 80%. Extra carbon dioxide additions and higher yeast extract concentrations were required to increase the MCFA production rate. More research related to substrates and (micro)nutrients in mixed culture continuous reactors needs to be performed to reduce yeast extract use in chain elongation.

High rate heptanoate production from propionate and ethanol using chain elongation

Heptanoate (or enanthate), a saturated mono-carboxylate with seven carbon atoms, is a commercially produced biochemical building block with versatile applications. Currently, heptanoate is mainly derived from the oxidation of heptaldehyde, which can be obtained after pyrolysis of castor oil. The objective of this investigation was to achieve efficient high rate heptanoate production using a mixed culture chain elongation process based on propionate and ethanol. An efficient high rate heptanoate production using chain elongation could offer an alternative for heptanoate production from castor oil. The investigation was performed in an upflow anaerobic filter with a hydraulic retention time of 17 h. A heptanoate production rate of 4.5 g l(-1) d(-1) was achieved with a heptanoate concentration of 3.2 g l(-1). These results show sufficient potential to consider this approach as an alternative for heptanoate production from castor oil. Future research should make heptanoate production from propionate and ethanol more cost-effective.