Abstract
The bacterium Kluyvera georgiana MCC 3673 transfers electrons directly to the electrode for bio-electricity generation in microbial fuel cell (MFC). This could be due to the formation of biofilm on the surface of electrode or with through the extracellular appendages, or both. The role of extracellular appendages pili and flagella in exo-electron transfer mechanism was investigated. The expression level of the genes fli P and pil Q for pili and flagella, respectively, in K. georgiana MCC 3673 was compared in MFC and in shake flask. The transcript analysis was done by qRT-PCR at different times and conditions. The expression level of pil Q transcript in K. georgiana MCC 3673 showed over twofold higher expression during bio-electrogenic process, compared to the one inoculated in shake flask. Similarly, fli P had also showed similar kind of expression in MFC compared to that in shake flask. Higher level of pil Q and fli P transcripts were observed throughout bio-electrogenic process. The level of pil Q was found to be nearly fourfold higher in biofilm-forming cells forming compared to the cells in suspension form. The obtained results suggest that flagella have a role in movement of bacterium towards electrode for donating the electron in absence of oxygen, and pili aiding in adhering on the surface of electrode and forming biofilm. The cumulative effect of fli P and pil Q resulted in exo-electron transfer to the electrode and bio-electricity generation process. The open circuit potential (OCV) of + 0.7 V was produced with the maximum power density of 393 ± 14 mW/m2 in MFC.




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Barker CS, Meshcheryakova IV, Inoue T, Samatey FA (2014) Assembling flagella in Salmonella mutant strains producing a type III export apparatus without FliO. J Bacteriol 196:4001–4011. https://doi.org/10.1128/JB.02184-14
Baudler A, Schmidt I, Langner M et al (2015) Does it have to be carbon? Metal anodes in microbial fuel cells and related bioelectrochemical systems. Energy Environ Sci 8:2048–2055. https://doi.org/10.1039/C5EE00866B
Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622. https://doi.org/10.1373/clinchem.2008.112797
Cao Y, Mu H, Liu W et al (2019) Electricigens in the anode of microbial fuel cells: pure cultures versus mixed communities. Microb Cell Fact 18:1–14. https://doi.org/10.1186/s12934-019-1087-z
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159. https://doi.org/10.1016/0003-2697(87)90021-2
Gorby YA, Yanina S, McLean JS et al (2006) Electrically conductive bacterial nanowires produced by Shewanella oneidensis strain MR-1 and other microorganisms. Proc Natl Acad Sci 103:11358–11363
Li SL, Nealson KH (2015) Enriching distinctive microbial communities from marine sediments via an electrochemical-sulfide-oxidizing process on carbon electrodes. Front Microbiol 6:1–8. https://doi.org/10.3389/fmicb.2015.00111
Liu R, Ochman H (2007) Stepwise formation of the bacterial flagellar system. Proc Natl Acad Sci USA 104:7116–7121. https://doi.org/10.1073/pnas.0700266104
Liu X, Liu H, Zhang L et al (2018) Biological synthesis of high-conductive pili in aerobic bacterium Pseudomonas aeruginosa. Appl Microbiol Biotechnol 103:1535–1544. https://doi.org/10.1007/s00253-018-9484-5
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Logan BE, Hamelers B, Rozendal R et al (2006) Microbial fuel cells: methodology and technology. Environ Sci Technol 40:5181–5192. https://doi.org/10.1021/es0605016
Lovley DR (2006) Microbial fuel cells: novel microbial physiologies and engineering approaches. Curr Opin Biotechnol 17:327–332. https://doi.org/10.1016/j.copbio.2006.04.006
Mattick JS (2002) Type IV pili and twitching motility. Annu Rev Microbiol 56:289–314. https://doi.org/10.1146/annurev.micro.56.012302.160938
Moens S, Vanderleyden J (1996) Functions of bacterial flagella. Crit Rev Microbiol 22:67–100. https://doi.org/10.3109/10408419609106456
Poggio S, Abreu-Goodger C, Fabela S et al (2007) A complete set of flagellar genes acquired by horizontal transfer coexists with the endogenous flagellar system in Rhodobacter sphaeroides. J Bacteriol 189:3208–3216. https://doi.org/10.1128/JB.01681-06
Proft T, Baker EN (2009) Pili in Gram-negative and Gram-positive bacteria—structure, assembly and their role in disease. Cell Mol Life Sci 66:613–635. https://doi.org/10.1007/s00018-008-8477-4
Rabaey K, Boon N, Höfte M, Verstraete W (2005) Microbial phenazine production enhances electron transfer in biofuel cells. Environ Sci Technol 39:3401–3408. https://doi.org/10.1021/es048563o
Reguera G, McCarthy KD, Mehta T et al (2005) Extracellular electron transfer via microbial nanowires. Nature 435:1098–1101. https://doi.org/10.1038/nature03661
Richter K, Schicklberger M, Gescher J (2012) Dissimilatory reduction of extracellular electron acceptors in anaerobic respiration. Appl Environ Microbiol 78:913–921. https://doi.org/10.1128/AEM.06803-11
Schröder U (2007) Anodic electron transfer mechanisms in microbial fuel cells and their energy efficiency. Phys Chem Chem Phys 9:2619–2629. https://doi.org/10.1039/b703627m
Segura A, Duque E, Hurtado A, Ramos JL (2001) Mutations in genes involved in the flagellar export apparatus of the solvent-tolerant Pseudomonas putida DOT-T1E strain impair motility and lead to hypersensitivity to toluene shocks. J Bacteriol 183:4127–4133. https://doi.org/10.1128/JB.183.14.4127-4133.2001
Shi W, Sun H (2002) Type IV pilus-dependent motility and its possible role in bacterial pathogenesis. Infect Immun 70:1–4. https://doi.org/10.1128/IAI.70.1.1-4.2002
Shrestha PM, Rotaru AE (2014) Plugging in or going wireless: strategies for interspecies electron transfer. Front Microbiol 5:1–8. https://doi.org/10.3389/fmicb.2014.00237
Sonawane JM, Yadav A, Ghosh PC, Adeloju SB (2017) Recent advances in the development and utilization of modern anode materials for high performance microbial fuel cells. Biosens Bioelectron 90:558–576. https://doi.org/10.1016/j.bios.2016.10.014
Thapa BS, Chandra TS (2019) Kluyvera georgiana MCC 3673: a novel electrogen enriched in microbial fuel cell fed with oilseed cake. Curr Microbiol 76(5):650–657. https://doi.org/10.1007/s00284-019-01673-0
Thapa BS, Seetharaman S, Chetty R, Chandra TS (2019) Xerogel based catalyst for improved cathode performance in microbial fuel cells. Enzym Microb Technol 124:1–8. https://doi.org/10.1016/j.enzmictec.2019.01.007
Tonjum T, Collins RF, Ford RC et al (2002) Analysis of the PilQ secretin from neisseria meningitidis by transmission electron microscopy reveals a dodecameric quaternary structure. J Bacteriol 183:3825–3832. https://doi.org/10.1128/jb.183.13.3825-3832.2001
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Authors are thankful to the DST-FIST for providing real time PCR facility in the Department of Biotechnology, IIT Madras.
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BST and TSC planned the study, BST performed the experiments, BST and TSC analysed the data, and BST drafted the manuscript. TSC corrected the manuscript. Both the authors have agreed for communicating the present work with the journal 3 Biotech.
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Thapa, B.S., Chandra, T.S. Studies on expression levels of pil Q and fli P genes during bio-electrogenic process in Kluyvera georgiana MCC 3673. 3 Biotech 10, 73 (2020). https://doi.org/10.1007/s13205-020-2050-8
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DOI: https://doi.org/10.1007/s13205-020-2050-8

