maximum of two exponential random variables

production of ethanol and butanol) to the production of carboxylic acids and PDO (see Fig. Growth of Geobacter sulfurreducens with acetate in syntrophic cooperation with hydrogen-oxidizing anaerobic partners. According to the different hypothesises, this acetate was used by G. sulfurreducens according to the following equation (1): Pathways are considered to be favoured or disfavoured if more than 10% increase of decrease resp. 102, 334341 (2011). This progress was Extracellular electron transfer from cathode to microbes: application for biofuel production. Through such interaction, G. sulfurreducens triggered a significant metabolic shift for C. pasteurianum that enhanced the production of PDO at the expense of butanol and ethanol. For each sample, 6 qPCR replicates were performed to assess the standard error of measurement of the technique. A well described example is the IET existing between archaea and bacteria during methanogenesis through the diffusion of H2 or formate2,3. Use the Previous and Next buttons to navigate three slides at a time, or the slide dot buttons at the end to jump three slides at a time. GSU2029 to GSU2039 include genes for PilM, PilN, PilO and PilP, which are thought to be part of the assembly responsible for pilus protrusion and retraction in G. sulfurreducens [30] as they serve this function in other bacteria [27,29]. When a small inoculum of G. sulfurreducens was introduced into electrode-containing chambers, electrical current production was dependent upon oxidation of acetate to carbon dioxide and increased exponentially, indicating for the first time that electrode reduction supported the growth of this organism. Hereby biomass increase occured faster compared to experiment 4F0%. FOIA 2010 Nov 22;9:90. doi: 10.1186/1475-2859-9-90. 2017 Sep 5;199(19):e00340-17. 205, 205212 (2016). Oxidation of acetate through reactions of the citric acid cycle by Geobacter sulfurreducens in pure culture and in syntrophic coculture. Cell growth of G . The yield of oxygen reduced per mole of acetate YO2/S was determined to be 1.76 0.12 molO2 molS-1 which is in the same range as, but more precise than the value of 2.4 0.8 mol mol-1 determined by Lin et al. Environ. 6, 6283 (2015). Indeed, a proteomic analysis previously revealed that under long-term terminal electron acceptor limiting conditions, G. sulfurreducens DSM 12127 enhanced the synthesis of some of its membrane-associated proteins36. Complete DSMZ Media List. In these two replicates, PDO and butyrate yields significantly increased to 2418mmol.molglycerol1 (p=0.016) and 1056mmol.molglycerol1 (p=0.018) respectively, with regard to C. pasteurianum alone. : 4609 Culture medium link https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium826.pdf Temperatures Kind of temperature Temperature Ref. Concentration profiles of fumarate (closed, Fig 5. There was only cysteine as oxygen capturing chemical species present in the cultivation medium, but due to the overnight equilibration of reactors at aerobic conditions, cysteine should already be completely reduced. Cell dry weight (filled triangles), acetate concentration (open triangles) and dissolved oxygen concentration, Fig 5. Caccavo, F. et al. Geobacter sulfurreducens Caccavo et al. - BAA-2815 | ATCC Geobacter sulfurreducens is a Gram-negative, rod-shaped -proteobacterium found in subsurface environments [1]. Cell dry weight (filled squares), acetate concentration (open squares) and dissolved oxygen concentration (line) during experiment 40F0% (40 mM fumarate, 0% oxygen). Cell dry weight (filled circles), acetate concentration (open circles) and dissolved oxygen concentration (line) during experiment 4F3% (4 mM fumarate, 3% oxygen). Arch. also determined the same biomass yield per mole of acetate with either fumarate or oxygen as electron acceptor [9]. 4. Geobacter sulfurreducens could be cultured under acetate-limiting conditions with fumarate or Fe(III)-citrate as the electron acceptor at growth rates between 0.04 and 0.09 h 1. During growth, hydrogen was detected in very slight amounts (5 Pa). PMC If your changes are legitimate, changes will occur within the next BacDive update. Environ. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. official website and that any information you provide is encrypted Simple absolute quantification method correcting for quantitative PCR efficiency variations for microbial community samples. In all successful microaerobic cultivations, the DO concentration was around 0 mg L-1 during the time of cell growth and acetate consumption. The growth of G. sulfurreducens was simulated under different growth conditions, varying electron donors, electron acceptors, and nitrogen sources. Belousov, Yu. Which of the following statements isare true origin designed and coordinated the study and helped to draft the manuscript. In contrast, butanol (r=0.87, p=0.0002, n=8) and ethanol (r=0.95, p=0.0008, n=8) were disfavoured when G. sulfurreducens cells increased. 2020 Aug;14(8):2078-2089. doi: 10.1038/s41396-020-0673-8. Sci. Growth of Geobacter sulfurreducens with, Growth of Geobacter sulfurreducens with CO and fumarate. 2006 Jan;188(2):450-5. doi: 10.1128/JB.188.2.450-455.2006. The maximum biomass concentration achieved in experiment 4F3% was the same as in the anaerobic control experiment 40F0%. Examination of two years of community dynamics in an anaerobic bioreactor using fluorescence polymerase chain reaction (PCR) single-strand conformation polymorphism analysis. government site. By submitting a comment you agree to abide by our Terms and Community Guidelines. Lovley, D. R. et al. YX/S was at 0.552 0.014 gCDW gS-1 under anaerobic conditions (40F0%) and was similar for all cultivations except 4F1%, which yielded a slightly higher value of 0.63 0.06 gCDW gS-1. 100mL serum bottles sealed with butyl rubber stoppers and aluminum crimp caps were used with a working volume of 50mL under a N2 gas phase (>99.995%). To date, studies on electron flux between exoelectrogens such as Geobacter species and fermenters have always focused on the degradation of organic matter by the fermenters into simple carboxylic acids that could be readily converted by the exoelectrogens into electricity in microbial fuel cells, i.e. Shi, L. et al. Zhao, Z., Zhang, Y., Wang, L. & Quan, X. The NgCpast of C. pasteurianum was also lower (NgCpast=6.60.6, n=8) than the value of 9.30.8 obtained in the previous experiment, as a consequence of a higher inoculum concentration. -. Geobacter sulfurreducens wild type (PCA, ATCC 51573) was routinely cultivated in 10 ml nutrient broth (NB) medium under a N 2:CO 2 atmosphere (80:20, 15 ml headspace) with 10 mM acetate and 40 mM fumarate at 30 C and pH 6.8 as described previously (Caccavo et al., 1994; Coppi et al., 2001) (Table S1). Jo Philips - Assistant Professor - Aarhus University | LinkedIn 2005;5:41. doi: 10.1186/1471-2180-5-41. Choi, O., Kim, T., Woo, H. M. & Um, Y. Electricity-driven metabolic shift through direct electron uptake by electroactive heterotroph Clostridium pasteurianum . To sustain their growth and maintenance, microorganisms perform oxidative and reductive reactions inside their cells. Thus, proton motive force can be generated 3 times as efficiently using c-type cytochromes compared to the bd-type, resulting in a higher ATP yield [21,24]. sharing sensitive information, make sure youre on a federal Shrestha, P. M. et al. Transcriptome data of G. sulfurreducens grown under microaerobic conditions with 4 mM of fumarate and 1% of oxygen provided in the gas inlet as electron acceptors. Biotechnol. Technol., 104 (2012), pp. Geobacter bacteria efficiently oxidize acetate into electricity in bioelectrochemical systems, yet the range of fermentation products that support the growth of anode biofilms and electricity production has not been thoroughly investigated. Theses pre-cultures were centrifuged at 3600g for 1mn. CAS Instead of using Fe3+, these species were reported to be able to transfer their electrons to other species, such as denitrifying bacteria9,10 or methanogens11. The authors have declared that no competing interests exist. As this fermenter was able to uptake extracellular electrons from a cathode, it is not excluded that electrons provided by DIET with an exoelectrogen organism could also be consumed and lead to a similar metabolic shift. 2). In this context, the acetate consumed by G. sulfurreducens for its own growth could be assessed from qPCR data and was estimated around 1.10.4mM (n=2). Front. 60: 37523759 (1994) for further . 2022 Oct;11(5):e1322. Print 2017 Oct 1. Biotechnology Letters A high G. sulfurreducens growth positively correlated with an increase of PDO (r=0.95, p=0.0002, n=8), butyrate (r=0.91, p=0.0003, n=8) and acetate (r=0.93, p=0.0069, n=8). Overall, between 76 and 90% of total variance of the metabolite yields were explained by the growth of G. sulfurreducens. Environ. No growth should occur on nonselective media. 9, 13591363 (2009). PDF 826: Geobacter Sulfurreducens Medium - Dsmz -, Bengelsdorf FR, Straub M, Drre P. Bacterial synthesis gas (syngas) fermentation. Expression of these genes returned to the levels found under anaerobic conditions with the increase of oxygen to 3%. This suggests there is a limit of oxygen manageable by the cells, which must be in the range between 3 to 5% of oxygen in the gas inlet. For full functionality of this site, please enable JavaScript. When growing together with C. pasteurianum, G. sulfurreducens could have also used glycerol fermentation end-products as sole electron acceptors. Christina Elisabeth Anna Engel, The corresponding genes were, however, not expressed at higher levels under microaerobic conditions in the microarray analysis performed here. To achieve this, cultivations with altering amounts of oxygen were performed. This indicates that the use of oxygen as electron acceptor might indeed enable the cells to utilise acetate more efficiently, however only if the sOUR is low. The images or other third party material in this article are included in the articles Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. The authors declare no competing financial interests. 2012 Dec 1;40(6):1295-301. doi: 10.1042/BST20120099. government site. 2004 May;186(10):3022-8. doi: 10.1128/JB.186.10.3022-3028.2004. 2021 Oct 19;6(43):28859-28870. doi: 10.1021/acsomega.1c03805. Uptake of self-secreted flavins as bound cofactors for extracellular The bacterial growth of G. soli was determined by quantitative real-time PCR . The purpose of the study reported here was to identify potential candidates via whole-genome gene expression analysis and evaluate these candidates with genetic approaches. sulfurreducens under, MeSH However, this bacterium was later shown to not only tolerate exposure to oxygen but also to use it as terminal electron acceptor. Thus, only a qualitative comparison of Lin et al. Bethesda, MD 20894, Web Policies -, Bond DR, Lovley DR. Electricity production by Geobacter sulfurreducens attached to electrodes. Microbiology. During pure culture of G. sulfurreducens, no growth occurred as initial and final cell counts were strictly identical (3.31.9 106 and 3.31.2 106 cells.mL1 respectively, n=8 replicates), resulting in Ng<0.5 (twice the standard error of qPCR measurements). The increase in cell dry weight (CDW) occurred faster when oxygen was provided in comparison to the anaerobic experiment 4F0%. 2013;34:16391651. HPLC analysis were performed at a flow rate of 0.7mL/min on an Aminex HPX-87H, 3007.8mm (Bio-Rad) column at a temperature of 35C. Extra electrons seemed to trigger metabolic regulations in favor of PDO pathway as main electron dissipation pathway. Carboxydotrophic growth of Geobacter sulfurreducens - PubMed Medium/media recipe (s) required for preparing 826: - Medium 141. Before Jessen GL, Chen LX, Mori JF, Nelson TEC, Slater GF, Lindsay MBJ, Banfield JF, Warren LA. & Schink, B. Cysteine-mediated electron transfer in syntrophic acetate oxidation by cocultures of Geobacter sulfurreducens and Wolinella succinogenes . Microbiol. A. Front Microbiol. Like in the other experiments, the DO concentration remained at 0 mg L-1 for the entire cell growth time. J. Proteome Res. Unable to load your collection due to an error, Unable to load your delegates due to an error. designed and coordinated the study and helped to draft the manuscript. 1). The cytochrome c oxidase was formerly suspected to be the enzyme responsible for oxygen reduction in G. sulfurreducens [9]. Electron mass balances closed between 98.3 and 100.0%, meaning that all major metabolic products were quantified (see Fig. Correspondingly, the expression of genes encoding for enzymes involved in glutamate metabolism was likewise upregulated up to 3.6-fold, i.e. These results correspond well with results from Mouser et al., who found genes involved in motility to be expressed at lower levels in G. uraniireducens when exposed to 5% of oxygen [13]. G. sulfurreducens was cultivated in a minimal medium containing 10 mM acetate as carbon source and electron donor and 40 mM fumarate as electron acceptor as described previously [3840]. Kato, S. Biotechnological Aspects of Microbial Extracellular Electron Transfer. Error bars represent the standard deviation of the replicates. Geobacter sulfurreducens metabolism at different donor/acceptor ratios. PLOS ONE promises fair, rigorous peer review, Cooperative growth of Geobacter sulfurreducens and Clostridium pasteurianum with subsequent metabolic shift in glycerol fermentation. Microbiol. As carbon and energy source, G. sulfurreducens utilizes acetate [1]. ABSTRACT. This experiment and time-point were chosen as the highest sOUR was observed here. Choi, O. Rotaru, A.-E. et al. Arch Microbiol. Correspondence to Geobacter sulfurreducens sp. the genes for the ferritin-like domain containing proteins, GSU2193 and GSU2967, and for a universal stress protein, GSU0515) were upregulated up to 9-fold. A value of 0.23 log2(cells).mL1 was found. 2017 Sep 5;199(19):e00340-17. BMC Microbiol [Internet]. Biotechnol. In the presence of CO, reduction of NADP(+) was observed at a rate comparable to CO oxidation coupled to fumarate reduction in vivo. You are using a browser version with limited support for CSS. Benomar, S. et al. Geobacter sulfurreducens produce electrically conductive pili that function as nano-wires to promote electron transfer to insoluble electron acceptors such as Fe(III) oxide and electrodes [5, 6]. This site needs JavaScript to work properly. However, the corresponding gene was not differentially expressed under any of the conditions investigated here. Limitations of microbial iron reduction under extreme conditions In the case of experiment 4FSI, a final pyruvate concentration of 0.73 mM is obtained (S1A Fig). 43, 39533958 (2009). Interspecies Electron Transfer via Hydrogen and Formate Rather than Direct Electrical Connections in Cocultures of Pelobacter carbinolicus and Geobacter sulfurreducens . For all co-culture experiments and control experiments with G. sulfurreducens and C. pasteurianum cultivated alone, a minimal medium containing glycerol as a carbon and electron source for C. pasteurianum and acetate as an electron donor for G. sulfurreducens was used. The obtained microarray data were normalized using the Loess normalization [44] which makes sure that the intensity of the staining is uniformly distributed across all arrays also of different slides and analysed as described previously [42,43]. designed the study and carried out the experiments. Genome Scale Mutational Analysis of Geobacter sulfurreducens Reveals Distinct Molecular Mechanisms for Respiration and Sensing of Poised Electrodes versus Fe(III) Oxides. As protrusion and retraction of pili is an ATP intensive process, G. sulfurreducens might only follow the strategy of fleeing the zone of oxygen, when the concentration is adequately low and fleeing may actually result in finding an anaerobic zone. Microarray slides were analysed using the Microarray C Scanner (Agilent Technologies, Santa Clara, California, USA) as described previously [43]. Cell dry weight (filled circles), acetate concentration (open circles) and dissolved oxygen concentration, Fig 4. and Bacterial Populations during the Proliferation of L. pneumophila in a Cooling Tower Facility. Instead, a non-linear relationship was found between G. sulfurreducens growth (i.e. 1OQW: A) were taken from RSCB Protein Data aromatic amino acids at C-terminus have been shown with black Bank. 3). Butyrate production enhancement by Clostridium tyrobutyricum using electron mediators and a cathodic electron donor. Geobacter sulfurreducens could be cultured under acetate-limiting conditions with fumarate or Fe (III)-citrate as the electron acceptor at growth rates between 0.04 and 0.09 h 1. Alum Addition Triggers Hypoxia in an Engineered Pit Lake. 3). 27, 8895 (2014). The Pt is a common electrocatalyst in MFC, some studies have shown that Pt modified titanium electrode has good electrical properties, and pure titanium electrode is not suitable for MFC anode material. Still, the cc(b/o)c3 cytochrome oxidase was found to be more important for protection against oxidative stress, and this was explained by the higher ATP-yield resulting from oxygen reduction with this oxidase [21]. This corresponds well to the degradation of succinate via the TCA cycle. Genome-wide analysis of the RpoN regulon in Geobacter sulfurreducens Cord-Ruwisch, R., Lovley, D. R. & Schink, B. Bansal, R. et al. If extracellular electrons uptake is an ATP-consuming process, then forcing electron consumption could be an interesting strategy to reduce biomass and by extension sludge formation during fermentation processes, leading to a better carbon and electron recovery. ATCC51573Geobacter sulfurreducens_ The stoichiometry of this reaction is 1:1 and accordingly a molar succinate (Suc) to fumarate (Fum) yield (YSuc/Fum) of 0.972 0.025 molSuc molFum-1 is seen in cultivation 40F0%. In Pseudomonas aeruginosa, minor pilins are thought to form initiation complexes from which the protrusion of a new pilus can commence [29]. Cascade degradation of organic matters in brewery wastewater using a continuous stirred microbial electrochemical reactor and analysis of microbial communities. doi: 10.1128/JB.00340-17. 826: GEOBACTER SULFURREDUCENS MEDIUM NH4Cl 1.50 g Na2HPO4 0.60 g KCl 0.10 g Na-acetate 0.82 g Modified Wolin's mineral solution 10.00 ml NaHCO3 2.50 g Na2-fumarate 8.00 g Wolin's vitamin solution (10x) 1.00 ml Distilled water 1000.00 ml Dissolve ingredients (except bicarbonate, fumarate and vitamins), sparge medium with Error bars indicate standard deviation from biological triplicates. 7, 14721482 (2013). 1994;161:345351. Cell dry weight (filled circles),, Fig 3. J Mol Biol. Metabolic Efficiency of Geobacter sulfurreducens Growing - SpringerLink eCollection 2021 Nov 2. PLoS ONE 15(1): Citation: Engel CEA, Vorlnder D, Biedendieck R, Krull R, Dohnt K (2020) Quantification of microaerobic growth of Geobacter sulfurreducens. This indicates that other substrates were used by the cells as precursor for fumarate, to create more of the electron acceptor. A study by Daniel Bond and Derek Lovley in 2003 showed that because of G . Biotechnol. The acetate concentration in experiment 40F0% decreased in an exponential manner concomitant with the increase of the biomass concentration for 18 h. Similarly, a linear increase of the biomass concomitant with a linear decrease of the acetate concentration could be seen in experiment 4F3% from 2 to 24 h of the cultivation. Venkata Mohan, S., Velvizhi, G., Annie Modestra, J. Quantitative Real-Time polymerase chain reaction (qPCR) was used to follow G. sulfurreducens and C. pasteurianum growth. (BAA-2815), . One standard curve was generated from each assay by using 10-fold dilutions in sterilized water (Gibco by Life Technologies) of a target plasmid (Eurofins Genomics, Germany). For each strain, the later was determined using 12 samples from a unique pre-culture. Contacts between the two partners can be ensured by the establishment of a biofilm on a conductive material1,5 (e.g. Extractions were verified using Infinite 200 PRO NanoQuant (Tecan Group Ltd., Mnnedorf, Zwitzerland). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The https:// ensures that you are connecting to the doi: 10.1128/AEM.69.3.1548-1555.2003. To assess the role of the menaquinol oxidase and the other enzymes possibly involved in oxygen removal, the corresponding genes should be deleted in future experiments, followed by the characterization of growth of the resulting mutants under different concentrations of oxygen. 64, 22322236 (1998). Environ. A c-type cytochrome and a transcriptional regulator responsible for enhanced extracellular electron transfer in Geobacter sulfurreducens revealed by adaptive . Around this time, the DO concentration abruptly increased to approximately 1 mg L-1 in experiment 4F3%. 515-535. 16S rRNA gene: U13928. Adv Microb Physiol. Clipboard, Search History, and several other advanced features are temporarily unavailable. They observed a similar dependency of the ability of oxygen reduction by G. sulfurreducens on the provided oxygen concentration. From the samples taken, 6 were used for the qPCR analyses (cell quantification) and the other 6 were freeze-dried (measurement of mass of dried cells). Nanomaterials | Free Full-Text | Growth of Lactic Acid Bacteria on Gold doi: 10.1002/mbo3.1322. Then, electron exchanges with a partner such as C. pasteurianum would be promoted as the two species would be physically connected through both pili networks and the conductive material38. It may be that the amount of oxygen in the headspace of 5% or 10% provided by these authors [9] was already too high to allow for an investigation of the increased YX/S that was observed in this study for cultivation 4F1%. doi:10.1371/journal.pone.0215341, Editor: Vasu D. Appanna, Laurentian University, CANADA, Received: March 29, 2019; Accepted: December 30, 2019; Published: January 16, 2020. Geobacter sulfurreducens Can Grow with Oxygen as a Terminal Electron Under the hypothesis that interactions between G. sulfurreducens and C. pasteurianum are non-specific, continuous co-culture fermentations with electro-active and fermentative bacteria would provide many benefits such as: (i) recycling electrons from undesired fermentation end-products to promote the production more reduced compounds; (ii) avoiding the accumulation of inhibitors such as acetic acid; (iii) purifying the fermentation medium by removing undesired metabolites. Biomass concentration was determined by periodic sampling (1 mL, 2 h interval) and measurement of the optical density (Libra S11, Biochrom, Berlin, Germany) at 600 nm (OD600). Institute of Biochemical Engineering, Technische Universitt Braunschweig, Braunschweig, Germany, Other major metabolic products were 1,3-propanediol (PDO, 17613mmol.molglycerol1), ethanol (542mmol.molglycerol1) and butyrate (767mmol.molglycerol1). In the experiment 4F1%, DO concentrations remained at 0 mg L-1 for the entire cultivation time. For precultures medium was prepared in serum bottles (125 mL with 50 mL liquid volume, Fisher Scientific, Hampton, New Hampshire, USA) and gassed with N2 for a minimum of 20 min to obtain anaerobic conditions. DO sensors were calibrated by a two-point calibration using 21% oxygen and 0% oxygen (corresponding to 100% N2), respectively, in the gas inlet.
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