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Najjarzadeh, N., Krige, A., Pamidi, T. R., Johansson, Ö., Enman, J., Matsakas, L., . . . Christakopoulos, P. (2020). Numerical modeling and verification of a sonobioreactor and its application on two model microorganisms. PLoS ONE, 15(3), Article ID e0229738.
Open this publication in new window or tab >>Numerical modeling and verification of a sonobioreactor and its application on two model microorganisms
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2020 (English)In: PLoS ONE, E-ISSN 1932-6203, Vol. 15, no 3, article id e0229738Article in journal (Refereed) Published
Abstract [en]

Ultrasound has many uses, such as in medical imaging, monitoring of crystallization, characterization of emulsions and suspensions, and disruption of cell membranes in the food industry. It can also affect microbial cells by promoting or slowing their growth and increasing the production of some metabolites. However, the exact mechanism explaining the effect of ultrasound has not been identified yet. Most equipment employed to study the effect of ultrasound on microorganisms has been designed for other applications and then only slightly modified. This results in limited control over ultrasound frequency and input power, or pressure distribution in the reactor. The present study aimed to obtain a well-defined reactor by simulating the pressure distribution of a sonobioreactor. Specifically, we 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, and a lignocellulose-degrading fungus, Fusarium oxysporum. Sonication resulted in increased malate production by Gsulfurreducens, but no major effect on growth. In comparison, morphology and growth of Foxysporum were more sensitive to ultrasound intensity. Despite considerable morphological changes at 4 W input power, the growth rate was not adversely affected; however, at 12 W, growth was nearly halted. The above findings indicate that the novel sonobioreactor provides an effective tool for studying the impact of ultrasound on microorganisms.

Place, publisher, year, edition, pages
PLOS, 2020
National Category
Bioprocess Technology Fluid Mechanics and Acoustics
Research subject
Biochemical Process Engineering; Engineering Acoustics
Identifiers
urn:nbn:se:ltu:diva-78111 (URN)10.1371/journal.pone.0229738 (DOI)32160222 (PubMedID)2-s2.0-85081204531 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-03-26 (alebob)

Available from: 2020-03-19 Created: 2020-03-19 Last updated: 2020-03-26Bibliographically approved
Krige, A. (2019). Microbial Fuel cells, applications and biofilm characterization. (Licentiate dissertation). Luleå: Luleå University of Technology
Open this publication in new window or tab >>Microbial Fuel cells, applications and biofilm characterization
2019 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

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.

Place, publisher, year, edition, pages
Luleå: Luleå University of Technology, 2019. p. 32
Series
Licentiate thesis / Luleå University of Technology, ISSN 1402-1757
Keywords
MFC, Microbial fuel cell, Raman microscopy, BES, Geobacter sulfurreducens
National Category
Other Environmental Biotechnology Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-73938 (URN)978-91-7790-398-7 (ISBN)978-91-7790-399-4 (ISBN)
Presentation
2019-06-19, E632, Luleå University of Technology, E building, Luleå, 14:00 (English)
Opponent
Supervisors
Funder
Swedish Research Council, 2014-05906
Available from: 2019-05-15 Created: 2019-05-14 Last updated: 2019-06-05Bibliographically approved
Krige, A., Sjöblom, M., Ramser, K., Christakopoulos, P. & Rova, U. (2019). On-line Raman spectroscopic study of cytochromes’ redox state of biofilms in microbial fuel cells. Molecules, 24(3), Article ID 646.
Open this publication in new window or tab >>On-line Raman spectroscopic study of cytochromes’ redox state of biofilms in microbial fuel cells
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2019 (English)In: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 24, no 3, article id 646Article in journal (Refereed) Published
Abstract [en]

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. 

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
Cytochrome-C, Geobacter sulfurreducens, Microbial fuel cell, Omc, Raman spectroscopy
National Category
Bioprocess Technology Applied Mechanics
Research subject
Biochemical Process Engineering; Experimental Mechanics
Identifiers
urn:nbn:se:ltu:diva-73003 (URN)10.3390/molecules24030646 (DOI)000458934000270 ()30759821 (PubMedID)2-s2.0-85061525740 (Scopus ID)
Note

Validerad;2019;Nivå 2;2019-02-26 (svasva)

Available from: 2019-02-26 Created: 2019-02-26 Last updated: 2019-05-14Bibliographically approved
Sjöblom, M., Matsakas, L., Krige, A., Rova, U. & Christakopoulos, P. (2017). Direct electricity generation from sweet sorghum stalks and anaerobic sludge. Industrial crops and products (Print), 108, 505-511
Open this publication in new window or tab >>Direct electricity generation from sweet sorghum stalks and anaerobic sludge
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2017 (English)In: Industrial crops and products (Print), ISSN 0926-6690, E-ISSN 1872-633X, Vol. 108, p. 505-511Article in journal (Refereed) Published
Abstract [en]

Dried sweet sorghum stalks were valorized as a raw material for electricity generation in a two chamber microbial fuel cell using anaerobic sludge from a biogas plant as inoculum. The maximum voltage obtained on the sorghum stalks at an operating temperature of 35 °C was 546 mV with a maximum power- and current density of 131 mW/m2 and 543 mA/m2, respectively. The coulombic efficiency was 2.2%. Polarization data indicated that Ohmic resistances were dominant with an internal resistance of 182 Ω. The total electrical energy per gram of dried sorghum stalks was 165 J/g. Enzymatic treatment of the sorghum stalks did not improve the total electrical energy obtained. A metabolic study demonstrated that the sugars were quickly fermented to formate, acetate, propionate, lactate and butyrate with acetate and butyrate being the dominant acids during electricity generation

Place, publisher, year, edition, pages
Elsevier, 2017
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-65042 (URN)10.1016/j.indcrop.2017.06.062 (DOI)000412959800058 ()2-s2.0-85023637390 (Scopus ID)
Note

Validerad; 2017; Nivå 2; 2017-08-14 (andbra)

Available from: 2017-08-14 Created: 2017-08-14 Last updated: 2019-05-14Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-3386-701x

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