| 1. |
- Bajracharya, Suman, et al.
(författare)
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Advances in cathode designs and reactor configurations of microbial electrosynthesis systems to facilitate gas electro-fermentation
- 2022
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Ingår i: Bioresource Technology. - : Elsevier. - 0960-8524 .- 1873-2976. ; 354
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Tidskriftsartikel (refereegranskat)abstract
- In gas fermentation, a range of chemolithoautotrophs fix single-carbon (C1) gases (CO2 and CO) when H2 or other reductants are available. Microbial electrosynthesis (MES) enables CO2 reduction by generating H2 or reducing equivalents with the sole input of renewable electricity. A combined approach as gas electro-fermentation is attractive for the sustainable production of biofuels and biochemicals utilizing C1 gases. Various platform compounds such as acetate, butyrate, caproate, ethanol, butanol and bioplastics can be produced. However, technological challenges pertaining to the microbe-material interactions such as poor gas-liquid mass transfer, low biomass and biofilm coverage on cathode, low productivities still exist. We are presenting a review on latest developments in MES focusing on the configuration and design of cathodes that can address the challenges and support the gas electro-fermentation. Overall, the opportunities for advancing CO and CO2-based biochemicals and biofuels production in MES with suitable cathode/reactor design are prospected.
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| 2. |
- Bajracharya, Suman, et al.
(författare)
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Chapter 12 - Advances in gas fermentation processes
- 2022
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Ingår i: Current Developments in Biotechnology and Bioengineering. - : Elsevier. - 9780323911672 ; , s. 321-351
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Microbial metabolism enables the sustainable synthesis of fuels and chemicals from gaseous substrates (H2, CO, and CO2), thus drastically diminishing the carbon load in the atmosphere. Various value-added biochemicals and biofuels, such as acetate, methane, ethanol, butanol, butyrate, caproate, and bioplastics, have been produced during the conversion of syngas or H2/CO2, using a variety of microorganisms as biocatalysts. Gas fermentation processes using acetogenic and methanogenic organisms are being extensively investigated. This chapter provides an overview of microbial CO and CO2 conversion technology, with an emphasis on recent developments and integration with renewable electricity for the generation of H2 or other forms of electron donors. A discussion on technological challenges in gas fermentation addresses issues, such as poor mass transfer, low microbial biomass, and low productivity. It also presents possible solutions based on the latest advances in bioelectrochemical processes including microbial gas electrofermentation. Finally, the chapter includes a sustainability analysis of the process and includes a brief update on commercially established companies operating gas fermentation systems. Overall, an integrated approach combining gas fermentation and renewable electricity offers an opportunity for the development of CO and CO2- based biochemical and biofuel production at commercial scale.
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| 3. |
- Bajracharya, Suman, et al.
(författare)
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Dual cathode configuration and headspace gas recirculation for enhancing microbial electrosynthesis using Sporomusa ovata
- 2022
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Ingår i: Chemosphere. - : Elsevier. - 0045-6535 .- 1879-1298. ; 287, Part 3
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Tidskriftsartikel (refereegranskat)abstract
- High-rate production of acetate and other value-added products from the reduction of CO2 in microbial electrosynthesis (MES) using acetogens can be achieved with high reducing power where H2 appears as a key electron mediator. H2 evolution using metal cathodes can enhance the availability of H2 to support high-rate microbial reduction of CO2. Due to the low solubility of H2, the availability of H2 remains limited to the bacteria. In this study, we investigated the performances of Sporomusa ovata for CO2 reduction when dual cathodes were used together in an MES, one was regular carbon cathode, and the other was a titanium mesh that allows higher hydrogen evolution. The dual cathode configuration was investigated in two sets of MES, one set had the usual S. ovata inoculated graphite rod, and another set had a synthetic biofilm-imprinted carbon cloth. Additionally, the headspace gas in MES was recirculated to increase the H2 availability to the bacteria in suspension. High-rate CO2 reduction was observed at −0.9 V vs Ag/AgCl with dual cathode configuration as compared to single cathodes. High titers of acetate (up to ∼11 g/L) with maximum instantaneous rates of 0.68–0.7 g/L/d at −0.9 V vs Ag/AgCl were observed, which are higher than the production rates reported in literatures for S. ovata using MES with surface modified cathodes. A high H2 availability supported the high-rate acetate production from CO2 with diminished electricity input.
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| 4. |
- Bajracharya, Suman, et al.
(författare)
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Microbial Electrosynthesis Using 3D Bioprinting of Sporomusa ovata on Copper, Stainless-Steel, and Titanium Cathodes for CO2 Reduction
- 2023
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Ingår i: Fermentation. - : MDPI. - 2311-5637. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- Acetate can be produced from carbon dioxide (CO2) and electricity using bacteria at the cathode of microbial electrosynthesis (MES). This process relies on electrolytically-produced hydrogen (H2). However, the low solubility of H2 can limit the process. Using metal cathodes to generate H2 at a high rate can improve MES. Immobilizing bacteria on the metal cathode can further proliferate the H2 availability to the bacteria. In this study, we investigated the performances of 3D bioprinting of Sporomusa ovata on three metal meshes—copper (Cu), stainless steel (SS), and titanium (Ti), when used individually as a cathode in MES. Bacterial cells were immobilized on the metal using a 3D bioprinter with alginate hydrogel ink. The bioprinted Ti mesh exhibited higher acetate production (53 ± 19 g/m2/d) at −0.8 V vs. Ag/AgCl as compared to other metal cathodes. More than 9 g/L of acetate was achieved with bioprinted Ti, and the least amount was obtained with bioprinted Cu. Although all three metals are known for catalyzing H2 evolution, the lower biocompatibility and chemical stability of Cu hampered its performance. Stable and biocompatible Ti supported the bioprinted S. ovata effectively. Bioprinting of synthetic biofilm on H2-evolving metal cathodes can provide high-performing and robust biocathodes for further application of MES.
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| 5. |
- Krige, Adolf, et al.
(författare)
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3D bioprinting on cathodes in microbial electrosynthesis for increased acetate production rate using Sporomusa ovata
- 2021
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Ingår i: Journal of Environmental Chemical Engineering. - : Elsevier. - 2213-3437. ; 9:5
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Tidskriftsartikel (refereegranskat)abstract
- Microbial Electrosynthesis (MES) is emerging as a promising technology for the decarbonization of the economy. The use of CO2 as a feedstock, i.e. through carbon capture and utilization, can create attractive business opportunities for the production of 3rd generation biofuels. Achieving highly efficient electron transfer from the cathode to microbes is one of the main challenges hindering the development of MES reactors. It is, therefore, essential to improve biofilm growth for enhanced electron-transfer during bioconversion.In this study we show the first use of 3D bio-printing for microbial electrosynthesis systems, creating a “synthetic biofilm”, containing Sporomusa ovata. The synthetic biofilm greatly improves the acetate production rate while drastically decreasing start-up time. Using H-cell reactors, poised at -0.8 V vs Ag/AgCl, with a synthetic biofilm printed on a carbon cloth electrode, an average acetate productivity of 47±5.1 g⋅day-1⋅m-2 (0.31 ±0.55 g⋅L-1⋅day-1) with a maximum productivity of 104 g⋅day1⋅m-2 (0.68 g⋅L-1⋅day-1) was achieved. This is an order of magnitude higher than typical S. ovata production rates, and 2-3 fold higher than reactors using specialized cathodes. Start-up of MES reactors typically require days, whereas a high production rate was achieved almost directly after the start-up (±40 hours) using the synthetic biofilm. Cyclic voltammetry data showed H2 formation occurred at much higher potentials than in the control reactors, (approx. -550 mV vs. -950 mV for controls). Imaging showed that the synthetic biofilm allowed for dense growth of S. ovata cells at the cathode, increasing electron transfer efficiency and potentially improving the bio-catalyzation of H2.
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| 6. |
- Krige, Adolf, et al.
(författare)
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A New Approach for Evaluating Electron Transfer Dynamics by Using In Situ Resonance Raman Microscopy and Chronoamperometry in Conjunction with a Dynamic Model
- 2020
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Ingår i: Applied and Environmental Microbiology. - : American Society for Microbiology. - 0099-2240 .- 1098-5336. ; 86:20
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Tidskriftsartikel (refereegranskat)abstract
- Geobacter sulfurreducens is a good candidate as a chassis-organism due to its ability to form thick, conductive biofilms, enabling long distance extracellular electron transfer (EET). Due to the complexity of EET pathways in G. sulfurreducens, a dynamic approach is required to study genetically modified EET rates in the biofilm. By coupling on-line resonance Raman microscopy with chronoamperometry, we were able to observe the dynamic discharge response in the biofilm's cytochromes to an increase in anode voltage. Measuring the heme redox state alongside the current allows for the fitting of a dynamic model using the current response and a subsequent validation of the model via the value of a reduced cytochrome c Raman peak. The modelled reduced cytochromes closely fitted the Raman response data from the G. sulfurreducens wild-type strain, showing the oxidation of heme groups in cytochromes until achieving a new steady state. Furthermore, the use of a dynamic model also allows for the calculation of internal rates, such as acetate and NADH consumption rates. The Raman response of a mutant lacking OmcS showed a sharper initial rate than predicted, followed by an almost linear decrease of the reduced mediators. The increased initial rate could be attributed to an increase in biofilm conductivity, previously observed in biofilms lacking OmcS. One explanation for this is that OmcS acts as a conduit between cytochromes; therefore deleting the gene restricts the electron transfer rate to the extracellular matrix. This could, however, be modelled assuming a linear oxidation rate of intercellular mediators.IMPORTANCE Bioelectrochemical systems can fill a vast array of application niches, due to the control of redox reactions that it offers. Although native microorganisms are preferred for applications such as bioremediation, more control is required for applications such as biosensors or biocomputing. The development of a chassis organism, in which the EET is well defined and readily controllable, is therefore essential. The combined approach in this work offers a unique way of monitoring and describing the reaction kinetics of a G. sulfurreducens biofilm, as well as offering a dynamic model that can be used in conjunction with applications such as biosensors.
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| 7. |
- Krige, Adolf, 1990-, et al.
(författare)
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Design and implementation of a low cost bio-printer modification, allowing for switching between plastic and gel extrusion
- 2021
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Ingår i: HardwareX. - : Elsevier. - 2468-0672.
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Tidskriftsartikel (refereegranskat)abstract
- Due to the high cost of bioprinters they are not feasible for proof of concept experiments or educational purposes. Furthermore, the more affordable DIY methods all disable the plastic printing capability of the original printer. Here we present an affordable bio-printing modification that is easy to install and maintains the original capabilities of the printer. The modification used mostly 3D printed parts and is based on the popular, open-source Prusa i3 3D printer. The modifications are kept as simple as possible and uses standard slicing software, allowing for installation by less experienced builders. By using disposable syringes and easily sterilizable parts, an aseptic bioprinting setup can be achieved, depending on the environment. It also allows for 2 component printing as well as UV curing. The bio-printing and curing capabilities were shown by printing and curing an artificial biofilm of an electro-active bacteria, Geobacter sulfurreducens, onto a carbon-cloth electrode which was used in a microbial fuel cell.
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| 8. |
- Krige, Adolf, 1990-
(författare)
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Microbial Fuel cells, applications and biofilm characterization
- 2019
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Since the 1900’s it has been known that microorganisms are capable of generating electrical power through extracellular electron transfer by converting the energy found organic compounds (Potter, 1911). Microbial fuel cells (MFCs) has garnered more attention recently, and have shown promise in several applications, including wastewater treatment (Yakar et al., 2018), bioremediation (Rosenbaum & Franks, 2014), biosensors (ElMekawy et al., 2018) desalination (Zhang et al., 2018) and as an alternative renewable energy source in remote areas (Castro et al., 2014). In MFCs catalytic reactions of microorganisms oxidize an electron donor through extracellular electron transfer to the anode, under anaerobic conditions, with the cathode exposed to an electron acceptor, facilitating an electrical current (Zhuwei, Haoran & Tingyue, 2007; Lovley, 2006). For energy production in remote areas a low cost and easily accessible feed stock is required for the MFCs. Sweet sorghum is a drought tolerant feedstock with high biomass and sugar yields, good water-use efficiency, established production systems and the potential for genetic improvements. Because of these advantages sweet sorghum stalks were proposed as an attractive feedstock (Rooney et al., 2010; Matsakas & Christakopoulos, 2013). Dried sweet sorghum stalks were, therefore, tested as a raw material for power generation in a MFC, with anaerobic sludge from a biogas plant as inoculum (Sjöblom et al., 2017a).Using sorghum stalks the maximum voltage obtained was 546±10 mV, the maximum power and current density of 131±8 mW/m2 and 543±29 mA/m2 respectively and the coulombic efficiency was 2.2±0.5%. The Ohmic resistances were dominant, at an internal resistance of 182±17 Ω, calculated from polarization data. Furthermore, hydrolysis of the dried sorghum stalks did not improve the performance of the MFC but slightly increased the total energy per gram of substrate. During the MFC operation, the sugars were quickly fermented to formate, acetate, butyrate, lactate and propionate with acetate and butyrate being the key acids during electricity generation.Efficient electron transfer between the microorganisms and the electrodes is an essential aspect of bio-electrochemical systems such as microbial fuel cells. In order to design more efficient reactors and to modify microorganisms, for enhanced electricity production, understanding the mechanisms and dynamics of the electron transport chain is important. It has been found that outer membrane C-type cytochromes (OMCs) (including omcS and omcZ discussed in this study) play a key role in the electron transport chain of Geobacter sulfurreducens, a well-known, biofilm forming, electro-active microorganism (Millo et al., 2011; Lovley, 2008). It was found that Raman microscopy is capable of providing biochemical information, i.e., the redox state of c-type cytochromes (cyt-C) without damaging the microbial biofilm, allowing for in-situ observation.Raman microscopy was used to observe the oxidation state of OMCs in a suspended culture, as well as in a biofilm of an MFC. First, the oxidation state of the OMCs of suspended cultures from three G. sulfurreducens strains (PCA, KN400 and ΔpilA) was analyzed. It was found that the oxidation state can also be used as an indicator of the metabolic state of the cells, and it was confirmed that PilA, a structural pilin protein essential for long range electron transfer, is not required for external electron transfer. Furthermore, we designed a continuous, anaerobic MFC enabling in-situ Raman measurements of G. sulfurreducens biofilms during electricity generation, while poised using a potentiostat, in order to monitor and characterize the biofilm. Two strains were used, a wild strain, PCA, and a mutant, ΔOmcS. The cytochrome redox state, observed through the Raman spectra, could be altered by applying different poise voltages to the electrodes. This change was indirectly proportional to the modulation of current transferred from the cytochromes to the electrode. This change in Raman peak area was reproducible and reversible, indicating that the system could be used, in-situ, to analyze the oxidation state of proteins responsible for the electron transfer process and the kinetics thereof.
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| 9. |
- Krige, Adolf, et al.
(författare)
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On-line Raman spectroscopic study of cytochromes’ redox state of biofilms in microbial fuel cells
- 2019
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Ingår i: Molecules. - : MDPI. - 1431-5157 .- 1420-3049. ; 24:3
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Tidskriftsartikel (refereegranskat)abstract
- Bio-electrochemical systems such as microbial fuel cells and microbial electrosynthesis cells depend on efficient electron transfer between the microorganisms and the electrodes. Understanding the mechanisms and dynamics of the electron transfer is important in order to design more efficient reactors, as well as modifying microorganisms for enhanced electricity production. Geobacter are well known for their ability to form thick biofilms and transfer electrons to the surfaces of electrodes. Currently, there are not many “on-line” systems for monitoring the activity of the biofilm and the electron transfer process without harming the biofilm. Raman microscopy was shown to be capable of providing biochemical information, i.e., the redox state of C-type cytochromes, which is integral to external electron transfer, without harming the biofilm. In the current study, a custom 3D printed flow-through cuvette was used in order to analyze the oxidation state of the C-type cytochromes of suspended cultures of three Geobacter sulfurreducens strains (PCA, KN400 and ∆pilA). It was found that the oxidation state is a good indicator of the metabolic state of the cells. Furthermore, an anaerobic fluidic system enabling in situ Raman measurements was designed and applied successfully to monitor and characterize G. sulfurreducens biofilms during electricity generation, for both a wild strain, PCA, and a mutant, ∆S. The cytochrome redox state, monitored by the Raman peak areas, could be modulated by applying different poise voltages to the electrodes. This also correlated with the modulation of current transferred from the cytochromes to the electrode. The Raman peak area changed in a predictable and reversible manner, indicating that the system could be used for analyzing the oxidation state of the proteins responsible for the electron transfer process and the kinetics thereof in-situ.
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| 10. |
- Krige, Adolf, et al.
(författare)
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Role of Glycosyltransferase in the increased malate production in G. sulfurreducens when using sonobioreactor
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Ultrasound has been shown to affect microbial cells by promoting or slowing their growth and increasing the production of some metabolites. Using an optimized a sonotrode to match the bottle frequency and compared it to measured results to verify the accuracy of the simulation. The measured pressure distribution spectrum presented the same overall trend as the simulated spectrum. However, the peaks were much less intense, likely due to non-linear events such as the collapse of cavitation bubbles.To test the application of the sonobioreactor in biological systems, two biotechnologically interesting microorganisms were assessed: an electroactive bacterium, Geobacter sulfurreducens. Sonication resulted in increased malate production by G. sulfurreducens, but no major effect on growth.
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