Next Article in Journal
Methods for Fungicide Efficacy Screenings: Multiwell Testing Procedures for the Oomycetes Phytophthora infestans and Pythium ultimum
Next Article in Special Issue
Isolation and Characterisation of Electrogenic Bacteria from Mud Samples
Previous Article in Journal
Average Daily Gain in Lambs Weaned at 60 Days of Age Is Correlated with Rumen and Rectum Microbiota
Previous Article in Special Issue
Enterococcus faecalis NADH Peroxidase-Defective Mutants Stain Falsely in Colony Zymogram Assay for Extracellular Electron Transfer to Ferric Ions
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Microorganisms
Volume 11
Issue 2
10.3390/microorganisms11020349
microorganisms-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Polyphasic Characterization of Geotalea uranireducens NIT-SL11 Newly Isolated from a Complex of Sewage Sludge and Microbially Reduced Graphene Oxide

by
Li Xie
†,
Naoko Yoshida
*,† and
Lingyu Meng
Department of Civil Engineering, Nagoya Institute of Technology (Nitech), Nagoya 466-8555, Japan
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Microorganisms 2023, 11(2), 349; https://doi.org/10.3390/microorganisms11020349
Submission received: 4 January 2023 / Revised: 25 January 2023 / Accepted: 27 January 2023 / Published: 30 January 2023
(This article belongs to the Special Issue New Electrogenic Microbes)
Browse Figures
Review Reports Versions Notes

Abstract

:
Graphene oxide (GO), a chemically oxidized sheet of graphite, has been used as a conductive carbon carrier of microbes to boost various bioelectrochemical reactions. However, the types of microbes that can reduce GO have rarely been investigated. In this study, a strain of GO-reducing bacteria, named NIT-SL11, which was obtained from a hydrogel of microbially reduced GO and anaerobic sludge that converts sewage to electricity, was phylogenically identified as a novel strain of Geotalea uraniireducens. Considering the current lack of information on the electrogenic ability of the bacterium and its physicochemical and chemotaxonomic characteristics, the polyphasic characterization of the Geotalea uraniireducens strain NIT-SL11 was performed. NIT-SL11 utilized various organic acids, such as lactate, benzoate, and formate, as electron donors and exhibited respiration using GO, electrodes, fumarate, and malate. The strain contained C16:1ω7c and C16:0 as the major fatty acids and MK-8 and 9 as the major respiratory quinones. The complete genome of NIT-SL11 was 4.7 Mbp in size with a G+C content of 60.9%, and it encoded 80 putative c-type cytochromes and 23 type IV pili-related proteins. The possible extracellular electron transfer (EET) pathways of the strain were the porin–cytochrome (Pcc) EET pathway and type IV pili-based pathway.

1. Introduction

Bioelectrochemical systems, including microbial fuel cells (MFCs) and microbial electrolysis cells (MECs), and their applications in wastewater treatment have received much attention [1]. In MFC wastewater treatment, microorganisms degrade organic matter and transfer electrons to the anode; the electrons recovered in the anode are later utilized to reduce oxygen at the cathode. Thus, the anode is a critical factor that affects the energy recovery in bioelectrochemical systems, especially in the case of wastewater with a low chemical oxygen demand [2,3]. Preferably, the anode exhibits chemical stability, good affinity for microbes, and a large surface area [4,5,6]. Thus, various 3D anodes have been applied to facilitate current recovery [7,8,9,10].
The use of graphene oxide (GO), the oxidized form of graphene, has facilitated electricity production in several MFCs via microbial the reduction of non-conductive GO to conductive reduced GO (rGO) [11]. GO promotes the growth of selective exoelectrogens [12,13] by serving as the extracellular electron acceptor and self-aggregating into a 3D-conductive hydrogel that embeds exoelectrogens and results in significantly stabler energy production than is the case with graphite felt (GF) [14] and electrochemically oxidized GF [15]. Accordingly, hydrogels have been applied in the recovery of electricity from wastewater [14,16,17], including a relatively large-scale (100 L) swine wastewater treatment system utilizing MFCs [18]. To date, various microorganisms have been reported to reduce GO, including the following genera: Shewanella [11], Escherichia [18], Citrifermentans [12,19], and Desulfuromonas [13,20], as well as taxa found in natural microcosms [21]. However, exoelectrogens in the rGO complex converting sewage to electricity have yet to be isolated and characterized. In this study, an electrogenic bacterium, designated NIT-SL11, was successfully isolated from the rGO complex that converts sewage to electricity and is described in detail. NIT-SL11 is capable of growing under anaerobic conditions by oxidizing acetate coupled with Fe(III) reduction and is phylogenetically identified as a strain of Geotalea uraniireducens. The potential extracellular electron-transferring (EET) pathway is also proposed based on the genome sequence.

2. Materials and Methods

2.1. Isolation and Growth Conditions of NIT-SL11

Strain NIT-SL11 was isolated from an rGO complex anode utilized to recover electricity from municipal sewage wastewater, as previously described [14]. Microbes in the rGO anode were cultivated on an anaerobic agar plate supplemented with 5.0 mM acetate, 5.0 mM disodium anthraquinone-2,6-disulfonate (AQDS-Na), and 10% sludge extract. The agar plate was composed of DHB-CO3AAQ medium, which is based on a DHB-CO3 medium [22] with the following modifications: replacement of GO with 5.0 mM AQDS and supplementation with 0.5% agarose. After 7–14 days of incubation of the plate at 28 °C, electrochemically active colonies were visible due to orange halos, which is the color of the reduced form of AQDS. The colony culture was then purified by repeated agar-shake cultivation using DHB-CO3-AFY medium, which is a modified DHB-CO3 medium supplemented with 5.0 mM acetate, 5.0 mM fumarate, 0.1% yeast extract, and 0.1% yeast extract. The colony was collected from the agar culture, purified by repeated agar-shaking cultivation, and then re-cultivated in liquid DHB-CO3-AF medium, that is, DHB-CO3 medium supplemented with 5.0 mM acetate and 5.0 mM fumarate. Lastly, based on the uniformity of the microscopic morphology and 16S rRNA gene sequences, one of these liquid cultures was selected and further purified by repeating the agar cultivation step. The purified culture was then phylogenetically identified based on sequencing of the 16SrRNA gene amplified from the cell lysate [23] and named strain NIT-SL11. Strain NIT-SL11 was routinely cultured in liquid DHB-CO3-AF. In total, 7–14 days of anaerobic cultivation at 28 °C were sufficient to achieve full growth of the bacterial colony.

2.2. Morphological, Physiological, and Biochemical Analyses

The morphology of strain NIT-SL11 was observed under a field-emission scanning electron microscope (JSM-7800F; JEOL Ltd., Tokyo, Japan) operating at 1.0 kV [19], and its spore-forming ability and Gram-stainability were checked using optical microscopy, as previously described [24]. The salinity, temperature, and pH tolerance of NIT-SL11 were evaluated by measuring the growth of cells in DHB-CO3-AF medium. Salinity tolerance was examined by supplementing with 0 to 8% (w/v) of NaCl. The pH of the bicarbonate-free medium was adjusted to the range of 5.2 to 8.6 by the addition of sodium bicarbonate in the medium and CO2 in the headspace for the pH tolerance tests. The cell growth at different temperatures was tested in the range between 4 °C and 10 to 40 °C, with approximately 5 °C intervals.
The ability of strain NIT-SL11 to utilize an e− donor was determined by observing cell growth with the following substances in combination with reduction of 5 mM fumarate: 10 mM of formate, acetate, butyrate, lactate, pyruvate, succinate, propionate, malate, isobutyrate, caproate, benzoate, phenol, methanol, isopropanol, ethanol, butanol, glucose, fructose, and glycerol, and 0.5 g/L of peptone and yeast extract. Potential electron acceptors used by strain NIT-SL11 were assayed by observing cell growth in 10 mM each of fumarate, malate, sulfate, and thiosulfate; 5 mM of AQDS and nitrate; and 20 g/L of elemental sulfur with oxidation of 5 mM acetate. The production of electric current by the strain NIT-SL11 was evaluated via electrochemical cultivation using a graphite plate inoculated with NIT-SL11, as previously described [12].

2.3. Chemotaxonomic Analysis

The isoprenoid quinones and cellular fatty acid composition were investigated by Techno Suruga Laboratory Co., Ltd. (Shizuoka, Japan), as described in a previous study [24]. Isoprenoid quinones were extracted as previously described by Tamaoka et al. [25]. The analysis of cellular fatty acids was carried out using cells grown in liquid DHB-CO3-AF medium at 28 °C for 14 days, and the fatty acid profile was obtained according to the protocol of the Sherlock Microbial Identification System v6.0 (MIS, Newark, DE, USA) by accessing the TSBA6 database.

2.4. Genetic Characterization

The genomic DNA of strain NIT-SL11 was extracted as previously described [26,27]. Sequencing was performed using a combination of Illumina and Nanopore sequencing reads. A total of 1.8 M reads (1.02 GMbp) of paired-end reads and 0.37 M reads (1.76 GMbp) of single reads were subjected to error removal using Short Read Manager and assembled using Unicycler (v0.4.7, https://github.com/rrwick/Unicycler/releases/tag/v0.4.7, accessed on 18 October 2021). Gap sequences were determined in silico using Geno Finisher (v7.0, in silico biology, inc. Yokohama, Japan) [28]. Gene prediction and annotation of the complete genome of strainNIT-SL11 were performed using DFAST (https://dfast.ddbj.nig.ac.jp/, accessed on 11 October 2021) [29]. Gene comparison of NIT-SL11 and other Geobacter species was based on bidirectional best hits at 40% identity and 80% query coverage by SEED Viewer v2.0 (https://rast.nmpdr.org/rast.cgi, accessed on 18 October 2021) [30] and BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 26 October 2021) in NCBI database. The complete genome sequence of strain NIT-SL11 was deposited in DDBJ/GenBank under the BioProject number PRJDB15015 with accession number AP027151.
The full-length 16S rRNA gene sequence of strain NIT-SL11 was extracted from the complete genome data, and those for all publicly available Geobacter spp. were downloaded from the NCBI database. Phylogenetic analyses were performed using MEGA X (v8, Max Planck Institute of Biochemistry (MPIB), Planegg, Germany) (https://www.megasoftware.net/, accessed on 18 October 2022) based on the neighbor-joining method [31]. Statistical support for the branches of the phylogenetic trees was determined using bootstrap analysis based on 1000 re-samplings [32].

3. Results and Discussion

3.1. Isolation of NIT-SL11

The NIT-SL11 colonies were successfully cultivated following the method described above (Section 2.2). The cells of strain NIT-SL11 were found to be Gram-negative, non-spore-forming, rod-shaped, and approximately 0.4 µm in width and 1.4 µm in length (Figure 1A).
In the electrochemical cultivation, the colonies of strain NIT-SL11 grew and generated an orange-colored biofilm (Figure 1B) on a graphite electrode supplemented with acetate. Simultaneously, an electric current was rapidly generated and reached its maximum, 0.55 mA/cm2, on day 8 (Figure 1C). The current decreased over time but increased immediately after the addition of acetate to the medium. These results demonstrate that NIT-SL11 grew by coupling extracellular electron transfer with the electrode via acetate oxidation.

3.2. Phylogenetic Identification Based on 16S rRNA Sequencing

Strain NIT-SL11 was found to contain two 16S rRNA operons (rrn1 and 2), and the 16S rRNA gene-based phylogenetic tree revealed that the two rrns formed a cluster with Geotalea uraniireducens Rf4T (Figure 2). The 16S rRNA gene sequence similarity was 98.12% to 98.17% with respect to the other members of the genus Geotalea. Based on the cut-off values of 98.2–99.0% [33], and the 98.65% [34] similarity within the single species, the strain NIT-SL11 can be identified as a novel strain of G. uraniireducens. The genus Geotalea was recently proposed by Lovely et al. [35] by dividing Geobacter into four genera: Geobacter, Trichlorobacter, Citrifermentans, and Geotalea [36]. The genus Geotalea includes three species: G. uraniireducens [37] and G. daltonii [38], which were isolated from subsurface sediments, and G. toluenoxydans, which was isolated from the oil of a former coal-gasification site [39]. The isolation of a novel strain of G. uraniireducens from sewage wastewater indicated the widespread distribution of Geotalea in various environments. Geotalea species are Gram-negative, non-spore-forming rods, which are obligately anaerobic and typically able to oxidize acetate via coupling with fumarate reduction, although the potential for e- donor and acceptor utilization has not been investigated in this genus (Table 1). To the best of our knowledge, the present study is the first to demonstrate electricity production by a pure strain of Geotalea.

3.3. Physiological and Biochemical Characterization

Strain NIT-SL11 was found to tolerate temperatures in the range of 20 to 35 °C (optimum at 25–30 °C), pH 5.6–8.3 (optimum at 6.0–6.7), and NaCl concentrations of 0 to 2% (with an optimum at 0–1%). As shown in Table 1, consistent with most Geotalea strains, NIT-SL11 utilized acetate, benzoate, formate, and pyruvate as e− donors, with fumarate as the electron acceptor. Moreover, NIT-SL11 grew with the oxidization of lactate, peptone, pyruvate, and yeast extract, but not butanol, butyrate, caproate, ethanol, fructose, glucose, glycerol, isobutyrate, isopropanol, malate, methanol, phenol, propionate, and succinate. Strain NIT-SL11 was able to reduce fumarate and malate using acetate as an electron donor, whereas AQDS was not utilized as an electron acceptor. Additionally, NIT-SL11 was unable to reduce elemental sulfur, sulfate, and thiosulfate, similar to most Geotalea strains. None of the Geotalea strains were able to reduce nitrate levels. The cells of strain NIT-SL11 did not show any apparent movement on the slide, suggesting that strain NIT-SL11 was non-motile.

3.4. Chemotaxonomic Characterization

To the best of our knowledge, the major respiratory quinones and cellular fatty acids of G. uranireducens have not been reported. Therefore, such an analysis was performed in the present study using NIT-SL11. As shown in Table 2, the predominant quinones of strain NIT-SL11 were MK-8 (94%) and MK-9 (6.3%), and the major quinone was consistent with G. toluenoxydans TMJ1T. The predominant cellular fatty acids of strain NIT-SL11 were C16:1ω7c (39%) and C16:0 (32%), similar to those of G. daltonii FRC-32T and G. uraniireducens Rf4T, but not G. toluenoxydans TMJ1T, for which the main cellular fatty acids are C16 : 0 and C18 : 0.

3.5. General Genomic Features

An overview of the genome of strain NIT-SL11 and a comparison with other strains of the genus Geotalea are shown in Table 2. The genome size of strain NIT-SL11 was estimated to be 4.19 Mb, which approximates that of G. toluenoxydans TMJ1T and G. daltonii FRC-32T, whereas G. uraniireducens Rf4T showed a relatively larger genome size of approximately 5 Mb. The genome of NIT-SL11 encodes 3860 CDSs, 56 tRNAs, and 6 rRNAs. The G+C content of strain NIT-SL11 was calculated as 63.1%, which is relatively higher than that of other Geotalea strains (Table 2).
A genome map is shown in Figure 3. The KEGG category revealed that the highest number of genes were related to the metabolism of amino acids and their derivatives (200 genes), followed by protein metabolism (141 genes) and carbohydrate metabolism (135 genes). Regarding energy conversion, NIT-SL11 was found to have CDSs, which metabolize organic acids, such as formate, lactate, and pyruvate. The strain also showed a gene set for the complete TCA cycle, including CDSs, which metabolize organic acid intermediates (fumarate and malate) of the TCA cycle. NIT-SL11 also contains a full set of genes associated with glycolysis; however, the bacteria cannot utilize glucose because of the lack of glucose transporters. For nitrogen metabolism, NIT-SL11 only had CDSs for nitrogen fixation but not for the assimilation and dissimilation of nitrate reduction and denitrification. The sulfur metabolism pathway of NIT-SL11 was incomplete due to the absence of reductase-related genes for sulfur and thiosulfate. NIT-SL11 contained a full set of genes for assimilatory sulfate reduction; however, it lacked an extracellular sulfate transport system substrate-binding protein. These results are consistent with the inability of NIT-SL11 to use sulfur, sulfate, and thiosulfate as electron acceptors (Table 1).

3.6. Putative c-Type Cytochromes

NIT-SL11 was found to possess 80 putative c-type cytochromes (Table 3) via sequence screening, as previously described [20]. Based on PROSITE prediction [40], most c-type cytochromes were present in the periplasmic (19), extracellular (14), outer (9), and cytoplasmic membranes (5), whereas few cytochromes were present in the cytoplasm (3) and inner membrane (1). The number of heme-binding motifs varied from one to twenty-seven.
Seventy-three c-type cytochromes are homologs of those functionally identified in Geobacter sulfurreducens PCA, a well-characterized model strain of Geobacter genus. GURASL_06470 and 18290 are homologous to CbcA; GURASL_02110, 32780, 32770, 32730, and 22490 are homologous to CbcL, CbcM, CbcN, CbcR, and CbcX, respectively; and GURASL_ 03150 is homologous to ImcH. GURASL_06660 and 03230 are homologous to PpcA, and GURASL_03240 and 21440 are homologous to PpcC and PpcE, respectively. GURASL_08460 and 28660 are homologous to OmaB, and GURASL_28710, 28700, and 28650 are homologous to OmcB. These cytochromes are involved in the porin–cytochrome (Pcc) EET pathways that transfer electron across the cell envelope [41], indicating that the Pcc pathway may be one of the major EET pathways in strain NIT-SL11 (Figure 4). The Cbc complexes and ImcH are inner-membrane cytochromes that oxidize quinol in the cytoplasmic membrane and transfer the released electrons to the periplasmic PpcA homologs [42,43], which transfer the electrons acquired from the cytoplasm to the OmcB-based outer-membrane complex (ombB-omaB-omcB). The OmcB-based conduit transfers electrons through the lipid bilayer of the proteoliposomes [44,45]. It has been reported that, in the case of Geobacter sulfurreducens, CbcL is required for the reduction of electron acceptors with reduction potentials at or below −100 mV, and ImcH is necessary for the reduction of electron acceptors with reduction potentials above −100 mV [20]. Thus, there may also be two different pathways in NIT-SL11 grown on electrodes poised at different oxidizing potentials—the CbcL-dependent and ImcH-dependent pathways (Figure 4)—by which electrons are transported out of the inner-membrane quinone pool.
Strain NIT-SL11 also contains homologs of several other outer-membrane cytochromes in Geobacter sulfurreducens: GURASL_06720 and 17250 are homologous to OmcE, whereas GURASL_17360 is homologous to OmcS, which is suggested to transfer electrons to the T4P apparatus [46]; GURASL_32490, 32200, and 32150 are homologous to OmcG; GURASL_32160, 32170, 18110, and 32550 are homologous to OmcH; GURASL_26970 and 32480 are homologous to OmcM; and GURASL_27550, 17570, 15180, 32470, 32560, 06450, 19400, 07200, and 28530 are homologous to OmcF, I, K, N, P, Q, V, X, and Y, respectively. Most of these Omc-proteins have been reported to be involved in Fe(III) reduction of Geobacter species [47,48] and are upregulated during the growth of G. uraniireducens on Fe(III) oxides and/or Mn(IV) oxides [47]. GURASL_30840, 30810, 09840, 32800, 17430, and 17440 are homologs of ExtA, D, G, K, Q, and R, respectively. These genes belong to the outer-membrane conduit ext cluster, which aid electron transfer across the outer membrane to the bacterial surface [49]. Additionally, GURASL_22590 is homologous to the periplasmic electron transfer protein PgcA, which facilitates respiration to Fe(III) oxides but not to electrodes [50]. GURASL_ 35130 is a homolog of NrfA, which catalyzes the reduction of nitrite to ammonium via the dissimilatory nitrate reduction to ammonium (DNRA) pathway [51].

3.7. Type IV Pilus (T4P)-Related Genes

Strain NIT-T3 contains 23 CDSs encoding T4P (Table 4). T4P are filamentous polymers of pilin monomers that undergo dynamic rapid polymerization and depolymerization from a pool of pilins [52]. The NIT-SL11 genome contained a full set of genes encoding T4P: one major pilin PilA, one minor pilin PilE, and other essential proteins for secretin (PilQ), alignment (PilM, O, and P), platform (PilC), retraction ATPases (PilT), and assembly ATPases (PilB). All CDSs were found to be well conserved in the Geobacter strains. The pilus polymer pilin protein PilA of G. sulfurreducens is an electrically conductive pilus that is known to be involved in the EET of solid electron acceptors [53]. Aromatic acids are key factors associated with conductivity. The aromatic acid content in the PilA homolog in NIT-SL11 was estimated to account for 8.45%, which is consistent with the range of PilA aromatic acid content in phylogenetically diverse bacteria (5.5–25.25%) [54]. This suggests that the polymer was conductive. Thus, T4P transfers electrons directly to oxides distant from cell surfaces, which may be the second EET pathway of strain NIT-SL11 (Figure 4).

3.8. Exoelectrogens That form the rGO Complex

The type of electrode material is a crucial determinant of the formation of biofilms and the performance of electron transfer at the cell–electrode interface, which affects electricity production in bioelectrochemical systems. Many attempts have been made to propose novel electrode materials (e.g., carbon brush, carbon fabric, and GO) for use in such systems, and improved performance has been confirmed [55]. As a promising electrode material, GO electrodes have shown stabler energy production than conventional electrode materials (GF), mainly due to the formation of a self-aggregated conductive hydrogel (rGO complex) by the interaction between exoelectrogens and GO [18]. However, not all exoelectrogens can interact with GO to form a hydrogel [12], and the mechanism of hydrogel formation is poorly understood. To date, it has been shown that two exopolysaccharide (EPS) components, alpha-polysaccharides and bGlcNAc polysaccharides, are key players in the hydrogel formation of Shewanella BC01 and CN32 [56], which can be generated during bacterial growth, are readily adsorbed by rGO, and are helpful in terms of the in situ gelling of the bacteria/GO complex [57,58]. However, the most-studied strain, S. oneidensis MR-1, cannot form a hydrogel due to the lack of relevant genes [58]. On the other hand, although constituting another representative exoelectrogen, studies regarding hydrogel formation by Geobacter are limited. These genes are not coded in only two of the known hydrogel-forming Geobacter species, C. bremensis R4 [19] and G. NIT-SL11, which indicates that Geobacter and Shewanella differ in terms of the mechanism of hydrogel formation. Given that Geobacter is more predominant in MFC for wastewater treatment, the isolation of strain NIT-SL11 expands our understanding of the hydrogel formation mechanism of Geobacter. Further studies should focus on clarifying the functional genes and interaction between Geobacter and GO, which could form the basis for developing novel and effective electrode materials and would further advance the applications of MFC.

4. Conclusions

In this study, Geotalea uraniireducens NIT-SL11 was obtained from a hydrogel of microbially reduced GO and anaerobic sludge. The isolated strain was found to utilize various organic acids as electron donors and respired with GO, electrodes, fumarate, and malate. The analysis of the genome of NIT-SL11 suggests two possible extracellular electron transfer pathways. In the first possible pathway, porin–cytochrome (Pcc) EET pathways that start with the transfer of electrons from the inner membrane quinone pool to the membrane-associated cytochromes Cbcl or ImcH, which transfer these electrons further to periplasmic electron carrier PpcA homologs, and the PpcA homologs transfer the electrons from the cytoplasm to OmcB-based or Ext-cluster outer-membrane conduits, which, finally, transfer electrons through the outer membrane. In the second possible pathway, the aromatic amino acid-rich conductive pili transfer electrons directly to oxides distant from the cell surfaces.

Author Contributions

L.X.: data curation, data analysis, and writing; N.Y.: isolation, data curation, conceptualization, writing, supervision, funding acquisition; L.M.: data curation, data analysis, and writing. All authors have read and agreed to the published version of the manuscript.

Funding

This study was supported by funding from MEXT/JSPS KAKENHI (grant numbers 18K18876 and 22H01625) and the JSPS Joint Research Program with the NSFC. Genome analysis was supported by MEXT KAKENHI (No. 221S0002).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available upon request from the corresponding author.

Acknowledgments

We would like to thank Tomomi Suzuki and Asuka Akita for their technical support in cultivation and genomic experiments. We would also like to thank Kyou Ikeru for her support in disposing of cytochrome information.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Logan, B.E.; Rabaey, K. Conversion of Wastes into Bioelectricity and Chemicals by Using Microbial Electrochemical. Science 2012, 337, 686–690. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  2. Sugioka, M.; Yoshida, N.; Yamane, T.; Kakihana, Y.; Higa, M.; Matsumura, T.; Sakoda, M.; Iida, K. Long-Term Evaluation of an Air-Cathode Microbial Fuel Cell with an Anion Exchange Membrane in a 226L Wastewater Treatment Reactor. Environ. Res. 2022, 205, 112416. [Google Scholar] [CrossRef] [PubMed]
  3. Yamane, T.; Yoshida, N.; Sugioka, M. Estimation of Total Energy Requirement for Sewage Treatment by a Microbial Fuel Cell with a One-Meter Air-Cathode Assuming Michaelis–Menten COD Degradation. RSC Adv. 2021, 11, 20036–20045. [Google Scholar] [CrossRef]
  4. Chaudhuri, S.K.; Lovley, D.R. Electricity Generation by Direct Oxidation of Glucose in Mediatorless Microbial Fuel Cells. Nat. Biotechnol. 2003, 21, 1229–1232. [Google Scholar] [CrossRef] [PubMed]
  5. Wei, J.; Liang, P.; Huang, X. Recent Progress in Electrodes for Microbial Fuel Cells. Bioresour. Technol. 2011, 102, 9335–9344. [Google Scholar] [CrossRef]
  6. Nagahashi, W.; Yoshida, N. Comparative Evaluation of Fibrous Artificial Carbons and Bamboo Charcoal in Terms of Recovery of Current from Sewage Wastewater. J. Gen. Appl. Microbiol. 2021, 67, 248–255. [Google Scholar] [CrossRef]
  7. Chen, S.; Liu, Q.; He, G.; Zhou, Y.; Hanif, M.; Peng, X.; Wang, S.; Hou, H. Reticulated Carbon Foam Derived from a Sponge-like Natural Product as a High-Performance Anode in Microbial Fuel Cells. J. Mater. Chem. 2012, 22, 18609–18613. [Google Scholar] [CrossRef]
  8. Chen, S.; He, G.; Hu, X.; Xie, M.; Wang, S.; Zeng, D.; Hou, H.; Schröder, U. A Three-Dimensionally Ordered Macroporous Carbon Derived from a Natural Resource as Anode for Microbial Bioelectrochemical Systems. ChemSusChem 2012, 5, 1059–1063. [Google Scholar] [CrossRef]
  9. Feng, Q.; Wang, Y.; Wang, T.; Zheng, H.; Chu, L.; Zhang, C.; Chen, H.; Kong, X.; Xing, X.H. Effects of Packing Rates of Cubic-Shaped Polyurethane Foam Carriers on the Microbial Community and the Removal of Organics and Nitrogen in Moving Bed Biofilm Reactors. Bioresour. Technol. 2012, 117, 201–207. [Google Scholar] [CrossRef]
  10. Chen, S.; He, G.; Liu, Q.; Harnisch, F.; Zhou, Y.; Chen, Y.; Hanif, M.; Wang, S.; Peng, X.; Hou, H.; et al. Layered Corrugated Electrode Macrostructures Boost Microbial Bioelectrocatalysis. Energy Environ. Sci. 2012, 5, 9769–9772. [Google Scholar] [CrossRef]
  11. Salas, E.C.; Sun, Z.; Lüttge, A.; Tour, J.M. Reduction of Graphene Oxide via Bacterial Respiration. ACS Nano 2010, 4, 4852–4856. [Google Scholar] [CrossRef]
  12. Yoshida, N.; Miyata, Y.; Doi, K.; Goto, Y.; Nagao, Y.; Tero, R.; Hiraishi, A. Graphene Oxide-Dependent Growth and Self-Aggregation into a Hydrogel Complex of Exoelectrogenic Bacteria. Sci. Rep. 2016, 6, 21867. [Google Scholar] [CrossRef]
  13. Yoshida, N.; Goto, Y.; Miyata, Y.; Thakur, V.K. Selective Growth of and Electricity Production by Marine Exoelectrogenic Bacteria in Self-Aggregated Hydrogel of Microbially Reduced Graphene Oxide. C 2016, 2016, 15. [Google Scholar] [CrossRef] [Green Version]
  14. Yoshida, N.; Miyata, Y.; Mugita, A.; Iida, K. Electricity Recovery from Municipal Sewage Wastewater Using a Hydrogel Complex Composed of Microbially Reduced Graphene Oxide and Sludge. Materials 2016, 9, 742. [Google Scholar] [CrossRef] [Green Version]
  15. Yoshida, N.; Miyata, Y.; Iida, K. Current Recovery from Sewage Wastewater Using Electrochemically Oxidized Graphite Felt. RSC Adv. 2019, 9, 39348–39354. [Google Scholar] [CrossRef] [Green Version]
  16. Goto, Y.; Yoshida, N. Preliminary Evaluation of a Microbial Fuel Cell Treating Artificial Dialysis Wastewater Using Graphene Oxide. AIP Conf. Proc. 2016, 1709, 020007. [Google Scholar] [CrossRef] [Green Version]
  17. Goto, Y.; Yoshida, N. Microbially Reduced Graphene Oxide Shows Efficient Electricity Ecovery from Artificial Dialysis Wastewater. J. Gen. Appl. Microbiol. 2017, 63, 165–171. [Google Scholar] [CrossRef] [Green Version]
  18. Goto, Y.; Yoshida, N. Scaling up Microbial Fuel Cells for Treating. Water 2019, 11, 1803. [Google Scholar] [CrossRef] [Green Version]
  19. Meng, L.; Xie, L.; Hirose, Y.; Nishiuchi, T.; Yoshida, N. Reduced Graphene Oxide Increases Cells with Enlarged Outer Membrane of Citrifermentans Bremense and Exopolysaccharides Secretion. Biosens. Bioelectron. 2022, 218, 114754. [Google Scholar] [CrossRef]
  20. Xie, L.; Yoshida, N.; Ishii, S.; Meng, L. Isolation and Polyphasic Characterization of Desulfuromonas Versatilis Sp. Nov., an Electrogenic Bacteria Capable of Versatile Metabolism Isolated from a Graphene Oxide-Reducing Enrichment Culture. Microorganisms 2021, 9, 1953. [Google Scholar] [CrossRef]
  21. Tanizawa, Y.; Okamoto, Y.; Tsuzuki, K.; Nagao, Y.; Yoshida, N.; Tero, R.; Iwasa, S.; Hiraishi, A.; Suda, Y.; Takikawa, H.; et al. Microorganism Mediated Synthesis of Reduced Graphene Oxide Films. In Journal of Physics: Conference Series, Proceedings of the Asia-Pacific Interdisciplinary Research Conference 2011 (AP-IRC 2011), Aichi, Japan, 17–18 November 2011; Institute of Physics Publishing: London, UK, 2012; Volume 352. [Google Scholar]
  22. Tomita, R.; Yoshida, N.; Meng, L. Formate: A Promising Electron Donor to Enhance Trichloroethene-to-Ethene Dechlorination in Dehalococcoides-Augmented Groundwater Ecosystems with Minimal Bacterial Growth. Chemosphere 2022, 307, 136080. [Google Scholar] [CrossRef] [PubMed]
  23. Yoshida, N.; Asahi, K.; Sakakibara, Y.; Miyake, K.; Katayama, A. Isolation and Quantitative Detection of Tetrachloroethene (PCE)-Dechlorinating Bacteria in Unsaturated Subsurface Soils Contaminated with Chloroethenes. J. Biosci. Bioeng. 2007, 104, 91–97. [Google Scholar] [CrossRef] [PubMed]
  24. Ismaeil, M.; Yoshida, N.; Katayama, A. Bacteroides Sedimenti Sp. Nov., Isolated from a Chloroethenes-Dechlorinating Consortium Enriched from River Sediment. J. Microbiol. 2018, 56, 619–627. [Google Scholar] [CrossRef] [PubMed]
  25. Tamaoka, J.; Katayama-Fujimura, Y.; Kuraishi, H. Analysis of Bacterial Menaquinone Mixtures by High Performance Liquid Chromatography. J. Appl. Bacteriol. 1983, 54, 31–36. [Google Scholar] [CrossRef]
  26. Ismaeil, M.; Yoshida, N.; Katayama, A. Identification of Multiple Dehalogenase Genes Involved in Tetrachloroethene-to-Ethene Dechlorination in a Dehalococcoides -Dominated Enrichment Culture. Biomed. Res. Int. 2017, 2017, 9191086. [Google Scholar] [CrossRef] [Green Version]
  27. He, F.E. Coli Genomic DNA Extraction. Bio. Protoc. 2011, 1, 4–6. [Google Scholar] [CrossRef]
  28. Ohtsubo, Y.; Maruyama, F.; Mitsui, H.; Nagata, Y.; Tsuda, M. Complete Genome Sequence of Acidovorax Sp. Strain KKS102, a Polychlorinated-Biphenyl Degrader. J. Bacteriol. 2012, 194, 6970–6971. [Google Scholar] [CrossRef] [Green Version]
  29. Tanizawa, Y.; Fujisawa, T.; Nakamura, Y. DFAST: A Flexible Prokaryotic Genome Annotation Pipeline for Faster Genome Publication. Bioinformatics 2018, 34, 1037–1039. [Google Scholar] [CrossRef] [Green Version]
  30. Overbeek, R.; Olson, R.; Pusch, G.D.; Olsen, G.J.; Davis, J.J.; Disz, T.; Edwards, R.A.; Gerdes, S.; Parrello, B.; Shukla, M.; et al. The SEED and the Rapid Annotation of Microbial Genomes Using Subsystems Technology (RAST). Nucleic Acids Res. 2014, 42, 206–214. [Google Scholar] [CrossRef]
  31. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol. Biol. Evol. 2018, 35, 1547–1549. [Google Scholar] [CrossRef]
  32. Felsenstein, J. Confidence Limits on Phylogenies: An Approach Using the Bootstrap. Evolution 1985, 39, 783–791. [Google Scholar] [CrossRef]
  33. Meier-Kolthoff, J.P.; Göker, M.; Spröer, C.; Klenk, H.P. When Should a DDH Experiment Be Mandatory in Microbial Taxonomy? Arch. Microbiol. 2013, 195, 413–418. [Google Scholar] [CrossRef] [PubMed]
  34. Kim, M.; Oh, H.S.; Park, S.C.; Chun, J. Towards a Taxonomic Coherence between Average Nucleotide Identity and 16S RRNA Gene Sequence Similarity for Species Demarcation of Prokaryotes. Int. J. Syst. Evol. Microbiol. 2014, 64, 346–351. [Google Scholar] [CrossRef]
  35. Lovley, D.R.; Giovannoni, S.J.; White, D.C.; Champine, J.E.; Phillips, E.J.P.; Gorby, Y.A.; Goodwin, S. Geobacter Metallireducens Gen. Nov. Sp. Nov., a Microorganism Capable of Coupling the Complete Oxidation of Organic Compounds to the Reduction of Iron and Other Metals. Arch. Microbiol. 1993, 159, 336–344. [Google Scholar] [CrossRef]
  36. Waite, D.W.; Chuvochina, M.; Pelikan, C.; Parks, D.H.; Yilmaz, P.; Wagner, M.; Loy, A.; Naganuma, T.; Nakai, R.; Whitman, W.B.; et al. Proposal to Reclassify the Proteobacterial Classes Deltaproteobacteria and Oligoflexia, and the Phylum Thermodesulfobacteria into Four Phyla Reflecting Major Functional Capabilities. Int. J. Syst. Evol. Microbiol. 2020, 70, 5972–6016. [Google Scholar] [CrossRef] [PubMed]
  37. Shelobolina, E.S.; Nevin, K.P.; Blakeney-Hayward, J.D.; Johnsen, C.V.; Plaia, T.W.; Krader, P.; Woodard, T.; Holmes, D.E.; VanPraagh, C.G.; Lovley, D.R. Geobacter Pickeringii Sp. Nov., Geobacter Argillaceus Sp. Nov. and Pelosinus Fermentans Gen. Nov., Sp. Nov., Isolated from Subsurface Kaolin Lenses. Int. J. Syst. Evol. Microbiol. 2007, 57, 126–135. [Google Scholar] [CrossRef] [PubMed]
  38. Prakash, O.; Gihring, T.M.; Dalton, D.D.; Chin, K.-J.; Green, S.J.; Akob, D.M.; Wanger, G.; Kostka, J.E. Geobacter Daltonii Sp. Nov., an Fe(III)- and Uranium(VI)-Reducing Bacterium Isolated from a Shallow Subsurface Exposed to Mixed Heavy Metal and Hydrocarbon Contamination. Int. J. Syst. Evol. Microbiol. 2010, 60, 546–553. [Google Scholar] [CrossRef] [PubMed]
  39. Kunapuli, U.; Jahn, M.K.; Lueders, T.; Geyer, R.; Heipieper, H.J.; Meckenstock, R.U. Desulfitobacterium Aromaticivorans Sp. Nov. and Geobacter Toluenoxydans Sp. Nov., Iron-Reducing Bacteria Capable of Anaerobic Degradation of Monoaromatic Hydrocarbons. Int. J. Syst. Evol. Microbiol. 2010, 60, 686–695. [Google Scholar] [CrossRef]
  40. Sigrist, C.J.A.; Cerutti, L.; Hulo, N.; Gattiker, A.; Falquet, L.; Pagni, M.; Bairoch, A.; Bucher, P. PROSITE: A Documented Database Using Patterns and Profiles as Motif Descriptors. Brief Bioinform 2002, 3, 265–274. [Google Scholar] [CrossRef]
  41. Jiang, Y.; Shi, M.; Shi, L. Molecular Underpinnings for Microbial Extracellular Electron Transfer during Biogeochemical Cycling of Earth Elements. Sci. China Life Sci. 2019, 62, 1275–1286. [Google Scholar] [CrossRef]
  42. Levar, C.E.; Chan, C.H.; Mehta-Kolte, M.G.; Bond, D.R. An Inner Membrane Cytochrome Required Only for Reduction of High Redox Potential Extracellular Electron Acceptors. mBio 2014, 5, e02034-14. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  43. Levar, C.E.; Hoffman, C.L.; Dunshee, A.J.; Toner, B.M.; Bond, D.R. Redox Potential as a Master Variable Controlling Pathways of Metal Reduction by Geobacter Sulfurreducens. ISME J. 2017, 11, 741–752. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  44. Liu, Y.; Fredrickson, J.K.; Zachara, J.M.; Shi, L. Direct Involvement of OmbB, OmaB, and OmcB Genes in Extracellular Reduction of Fe(III) by Geobacter Sulfurreducens PCA. Front. Microbiol. 2015, 6, 1075. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Liu, Y.; Wang, Z.; Liu, J.; Levar, C.; Edwards, M.J.; Babauta, J.T.; Kennedy, D.W.; Shi, Z.; Beyenal, H.; Bond, D.R.; et al. A Trans-Outer Membrane Porin-Cytochrome Protein Complex for Extracellular Electron Transfer by Geobacter Sulfurreducens PCA. Environ. Microbiol. Rep. 2014, 6, 776–785. [Google Scholar] [CrossRef] [Green Version]
  46. Shi, L.; Squier, T.C.; Zachara, J.M.; Fredrickson, J.K. Respiration of Metal (Hydr)Oxides by Shewanella and Geobacter: A Key Role for Multihaem c-Type Cytochromes. Mol. Microbiol. 2007, 65, 12–20. [Google Scholar] [CrossRef] [Green Version]
  47. Aklujkar, M.; Coppi, M.V.; Leang, C.; Kim, B.C.; Chavan, M.A.; Perpetua, L.A.; Giloteaux, L.; Liu, A.; Holmes, D.E. Proteins Involved in Electron Transfer to Fe(III) and Mn(IV) Oxides by Geobacter Sulfurreducens and Geobacter Uraniireducens. Microbiology 2013, 159, 515–535. [Google Scholar] [CrossRef] [Green Version]
  48. Butler, J.E.; Young, N.D.; Lovley, D.R. Evolution of Electron Transfer out of the Cell: Comparative Genomics of Six Geobacter Genomes. BMC Genom. 2010, 11, 40. [Google Scholar] [CrossRef] [Green Version]
  49. Otero, F.J.; Chan, C.H.; Bond, D.R. Identification of Different Putative Outer Membrane Electron Conduits Necessary for Fe(III) Citrate, Fe(III) Oxide, Mn(IV) Oxide, or Electrode Reduction by Geobacter Sulfurreducens. J. Bacteriol. 2018, 200, 1–20. [Google Scholar] [CrossRef] [Green Version]
  50. Zacharoff, L.A.; Morrone, D.J.; Bond, D.R.; Emerson, D.; Bond, D.R. Geobacter Sulfurreducens Extracellular Multiheme Cytochrome PgcA Facilitates Respiration to Fe (III) Oxides But Not Electrodes. Front. Microbiol. 2017, 8, 2481. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  51. Campeciño, J.; Lagishetty, S.; Wawrzak, Z.; Alfaro, V.S.; Lehnert, N.; Reguera, G.; Hu, J.; Hegg, E.L. Cytochrome c Nitrite Reductase from the Bacterium Geobacter Lovleyi Represents a New NrfA Subclass. J. Biol. Chem. 2020, 295, 11455–11465. [Google Scholar] [CrossRef] [PubMed]
  52. Forest, K.T. Type IV Pili: Dynamics, Biophysics and Functional Consequences. Nat. Rev. Microbiol. 2019, 17, 429–440. [Google Scholar] [CrossRef]
  53. Lovley, D.R.; Walker, D.J.F.; Risgaard-petersen, N.; Lovley, D.R. Geobacter Protein Nanowires. Front. Microbiol. 2019, 10, 2078. [Google Scholar] [CrossRef] [Green Version]
  54. Walker, D.J.F.; Adhikari, R.Y.; Holmes, D.E.; Ward, J.E.; Woodard, T.L.; Nevin, K.P.; Lovley, D.R. Electrically Conductive Pili from Pilin Genes of Phylogenetically Diverse Microorganisms. ISME J. 2018, 12, 48–58. [Google Scholar] [CrossRef] [Green Version]
  55. Noori, M.T.; Vu, M.T.; Ali, R.B.; Min, B. Recent advances in cathode materials and configurations for upgrading methane in bioelectrochemical systems integrated with anaerobic digestion. Chem. Eng. J. 2020, 392, 123689. [Google Scholar] [CrossRef]
  56. Shen, L.; Jin, Z.; Wang, D.; Wang, Y.; Lu, Y. Enhance wastewater biological treatment through the bacteria induced graphene oxide hydrogel. Chemosphere 2018, 190, 201–210. [Google Scholar] [CrossRef]
  57. Chen, X.; Chen, B. Macroscopic and spectroscopic investigations of the adsorption of nitroaromatic compounds on graphene oxide, reduced graphene oxide, and graphene nanosheets. Environ. Sci. Technol. 2015, 49, 6181–6189. [Google Scholar] [CrossRef]
  58. Lee, B.M.; Hur, J. Adsorption Behavior of Extracellular Polymeric Substances on Graphene Materials Explored by Fluorescence Spectroscopy and Two-Dimensional Fourier Transform Infrared Correlation Spectroscopy. Environ. Sci. Technol. 2016, 50, 6181–6189. [Google Scholar] [CrossRef]
Microorganisms 11 00349 g001 550
Figure 1. Morphology and electrogenic properties of strain NIT-SL11. (A) Scanning electron microscopic image of strain NIT-SL11. (B) Biofilm generated on an electrode in an electrochemical culture. (C) Electric current production by strain NIT-SL11. The arrow in panel C indicates a spike in acetate addition.
Figure 1. Morphology and electrogenic properties of strain NIT-SL11. (A) Scanning electron microscopic image of strain NIT-SL11. (B) Biofilm generated on an electrode in an electrochemical culture. (C) Electric current production by strain NIT-SL11. The arrow in panel C indicates a spike in acetate addition.
Microorganisms 11 00349 g001
Microorganisms 11 00349 g002 550
Figure 2. Phylogenetic tree generated using 16S rRNA gene sequences of members of Geobacteraceae family. Yellow circles and green diamonds indicate bootstraps >80% and <60%, respectively. GenBank accession numbers are stated in parentheses; GO: the ability to respire graphene oxide; Current: ability to generate current.
Figure 2. Phylogenetic tree generated using 16S rRNA gene sequences of members of Geobacteraceae family. Yellow circles and green diamonds indicate bootstraps >80% and <60%, respectively. GenBank accession numbers are stated in parentheses; GO: the ability to respire graphene oxide; Current: ability to generate current.
Microorganisms 11 00349 g002
Microorganisms 11 00349 g003 550
Figure 3. Features of the complete genome of strain NIT-SL11. Circular representation of the genome was generated using Tbtools-Ⅱ (v1.106, https://github.com/CJ-Chen/TBtools-Manual). Rings numbered from the outside to inside are: 1, location of tRNA (red), Transfer messenger RNA (blue), and rRNA (green); 2, gene density; 3, c-type cytochromes (black) and type IV pili (red); 4, G+C skew (red, positive; green, negative); 5, protein-coding sequences colored based on KEGG category; 6, G+C content; and 7, links showing repetitive sequence ≥95% identity (pink, >500 bp; purple, >2 kbp). KEGG, Kyoto Encyclopedia of Genes and Genomes.
Figure 3. Features of the complete genome of strain NIT-SL11. Circular representation of the genome was generated using Tbtools-Ⅱ (v1.106, https://github.com/CJ-Chen/TBtools-Manual). Rings numbered from the outside to inside are: 1, location of tRNA (red), Transfer messenger RNA (blue), and rRNA (green); 2, gene density; 3, c-type cytochromes (black) and type IV pili (red); 4, G+C skew (red, positive; green, negative); 5, protein-coding sequences colored based on KEGG category; 6, G+C content; and 7, links showing repetitive sequence ≥95% identity (pink, >500 bp; purple, >2 kbp). KEGG, Kyoto Encyclopedia of Genes and Genomes.
Microorganisms 11 00349 g003
Microorganisms 11 00349 g004 550
Figure 4. Proposed EET pathway of strain NIT-SL11. OM: outer membrane; CM: cytoplasmic membrane.
Figure 4. Proposed EET pathway of strain NIT-SL11. OM: outer membrane; CM: cytoplasmic membrane.
Microorganisms 11 00349 g004
Table
Table 1. Comparison of morphological and physiological properties of strain NIT-SL11 and recognized species of the genus Geotalea.
Table 1. Comparison of morphological and physiological properties of strain NIT-SL11 and recognized species of the genus Geotalea.
Strain NIT-SL11G. toluenoxydans TMJ1TG. daltonii FRC-32TG. uraniireducens Rf4T
Size (µm)0.4 × 1.40.4 × 2.1–3.80.3–0.5 × 1.0–1.50.5–0.6 × 1.2–2.0
Motility−−ND+
Optimum Temp. (°C)25–3025–323032
Optimum pH6.0–.76.6–7.06.7–7.36.5–7.0
NaCl (%)0–2ND0–0.7ND
e− donor
Butanol−ND+−
Butyrate−++−
Caproate−NDNDND
Ethanol−NDND+
Fructose−NDNDND
Glucose−NDNDND
Glycerol−NDNDND
Isobutyrate−NDNDND
Isopropanol−NDNDND
Malate−−NDND
Methanol−NDND−
Phenol−+NDND
Propionate−+−−
Succinate−−−−
Acetate++ND+
Benzoate+++ND
Formate+++−
Lactate+−−+
Peptone+NDNDND
Pyruvate++ND+
YE+NDNDND
H2ND−−−
e− acceptors
Nitrate−−−−
Sulfate−−ND−
Thiosulfate−−ND−
AQDS−NDND+
S0−−+−
Fumarate++++
Malate+ND++
GO+NDNDND
Electrode+NDNDND
Fe(III)ND++−
Mn(Ⅳ)ND−NDND
ND: data not obtained. T: typical strain.
Table
Table 2. Comparison of chemotaxonomic and genomic properties of strain NIT-SL11 and recognized species of the genus Geotalea.
Table 2. Comparison of chemotaxonomic and genomic properties of strain NIT-SL11 and recognized species of the genus Geotalea.
Strain NIT-SL11G. toluenoxydans TMJ1TG. daltonii FRC-32TG. uraniireducens Rf4T
Size (Mb)4.24.24.35.1
GC (%)60545354
Total predicted genes3923−38524591
CDSs3860−37454457
rRNA6−66
tRNA56−4949
CytC80−7369
Menaquinone8, 98−−
Fatty acids >40%−C16:0−−
30–40%C16:1ω7c , C16:0−C16:1ω7c , C16:0C16:1ω7c
20–30%−C18:0−C16:0
10–20%−−ios-C15:0ios-C15:0 ,C14:0
T: typical strain.
Table
Table 3. List of putative c-type cytochrome proteins present in the NIT-SL11 genome.
Table 3. List of putative c-type cytochrome proteins present in the NIT-SL11 genome.
TagLocalCXnCH123
n = 2n = 3n = 4
01590−3000105−4255
02110CM800cbcL,027436860125
03150CM610imcH,325910140861
03230−300ppcA,061214264121
03240PP300ppcC,036514264121
06040−300053314013909
06440CP201059115903656
06450OM1200omcQ,059221703655
06470PP700cbcA,059434032712
06660PP300ppcA,061214264121
06690PP920061514293840
06700OM800ctcB,061614303839
06720OM400omcE,061816853837
07200PP1200omcX,067008300641
07560−500extCF,272516851838
08460−800omaB,273816810988
08470−900omcB,273716820989
08960PP200imcG,153816851316
09830−500extCF,272516851838
09840EX1211extG,2724−1837
13150−100128422562997
15110−2700221021703135
15170OM1002204−−
15180PP1000omcK,220316852291
15200−800cytT,229921750672
17180CM100coxB,0222−0414
17250EX400omcE,061816853837
17260EX8002076−−
17360−600omcS,250331661989
17430PP2600extQ,249531601995
17440PP1600extR,249431591996
17570PP910omcI,122829122291
17990PP200macA,046625791316
18080CM8002076−−
18110−1301omcH,288408311834
18290CM700cbcA,059434032712
19400PP1200omcV,199623792822
21440PP300ppcE,176014263843
21680PP100cycC,1740−2136
22490−500cbcX,1648−2381
22590EX200pgcA,176131762022
22840−310ctcD,1785−−
22850CP510178626411922
22860−400178726401921
25430IM100ccoP,2513−1913
25830−300321416853748
26520EX8002076−−
26970EX600omcM,2294−−
27550OM100omcF,2432−0331
28090EX1403omcN,289831322035
28530OM800omcY,220121753130
28650EX1000omcB,273718610989
28660−800omaB,273816810988
28700EX1200omcB,273716820994
28710−800omaB,273816810988
28780PP100petJ,2743−0331
29020OM1403omcN,289831322035
30810−500extD,2642−−
30820−401extCF,264316811838
30840PP1110extA,264516850641
31410OM500ctcC,28010309−
32150−2004omcG,288231323427
32160−1904omcH,288331323427
32170−2403omcH,288416853428
32200−2105omcG,070216852035
32390OM2303omcN,289815133428
32470EX2003omcN,289815132035
32480EX1508omcM,289916850076
32490−2007omcG,070215133427
32550EX2313omcH,291216853428
32560EX400omcP,291314323837
32700−200292713594833
32730−400cbcR,293013480460
32770CP1000cbcN,293413440456
32780EX1200cbcM,29351343−
32800PP500extK,293713400447
34580PP90031371724−
34890−200276716854149
35130PP400nrfA,315431110665
37130−200333238184408
Microorganisms 11 00349 i001
1: Geobacter. sulfurreducens PCA1; 2: G. daltonii FRC-32; 3: G. uraniireducens Rf4. Tag indicates the locus tag of the coding sequence encoding c-type cytochromes in the genome. CM, cytoplasmic membrane; PP, periplasm; EX, extracellular; (−), unknown. The number and color scale for the closest protein in the strain indicate the locus tag and amino acid identity (%), respectively.
Table
Table 4. List of T4P assembly-related genes in the NIT-SL11 genome.
Table 4. List of T4P assembly-related genes in the NIT-SL11 genome.
TagAnnotationAromatic Acid Mole %123
20580geopilin domain 1 protein pilA-N8.5GSU1496Geob3369Gura2677
20590geopilin domain 2 protein pilA-C8.8GSU1497−−
33430PilZ domain protein11.0GSU3028Geo0640Gura4082
28450PilZ domain protein9.7GSU1051Geob0928Gura0726
17690PilZ domain protein10.7GSU1240Geob0815Gura0044
36280PilZ domain protein8.6GSU0137−Gura3986
01060PilZ domain protein8.2GSU0078−−
37060PilZ domain protein10.6GSU0312Geob1410Gura3298
20560sensor histidine kinase PilS11.0−−−
20570sigma-54-dependent transcriptional response pilR8.3−−−
36190twitching motility pilus retraction protein pilT-18.6−−−
01730twitching motility pilus retraction protein pilT-38.5−−−
20540twitching motility pilus retraction protein pilT-47.4−−−
26020type IV pilus assembly lipoprotein PilP5.0GSU2029Geob3067Gura1813
26080type IV pilus assembly protein PilY110.2GSU1066Geob3067−
20530type IV pilus biogenesis ATPase PilB6.7−−−
26050type IV pilus biogenesis ATPase PilM6.8−−−
26040type IV pilus biogenesis ATPase PilM4.2GSU3069Geob3069Gura1811
26030type IV pilus biogenesis protein PilO0.0GSU2030Geob3068Gura1812
20550type IV pilus inner membrane protein PilC7.9−−−
26130type IV pilus minor pilin PilE10.2GSU3548−−
26010type IV pilus secretin lipoprotein PilQ5.8−−−
26160type IV prepilin-like proteins leader peptide pilD15.1GSU2043Geob3081Gura1794
Identity (%)20–3031–4041–5051–6061–7071–8081–9091–100
Tag indicates the locus tag of coding sequences encoding T4P-related proteins in the genome. The number and color scale for the closest protein in the strain indicate the locus tag and amino acid identity (%), respectively. 1: Geobacter sulfurreducens PCA1; 2. G. daltonii FRC-32; 3. G. uraniireducens Rf4.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Xie, L.; Yoshida, N.; Meng, L. Polyphasic Characterization of Geotalea uranireducens NIT-SL11 Newly Isolated from a Complex of Sewage Sludge and Microbially Reduced Graphene Oxide. Microorganisms 2023, 11, 349. https://doi.org/10.3390/microorganisms11020349

AMA Style

Xie L, Yoshida N, Meng L. Polyphasic Characterization of Geotalea uranireducens NIT-SL11 Newly Isolated from a Complex of Sewage Sludge and Microbially Reduced Graphene Oxide. Microorganisms. 2023; 11(2):349. https://doi.org/10.3390/microorganisms11020349

Chicago/Turabian Style

Xie, Li, Naoko Yoshida, and Lingyu Meng. 2023. "Polyphasic Characterization of Geotalea uranireducens NIT-SL11 Newly Isolated from a Complex of Sewage Sludge and Microbially Reduced Graphene Oxide" Microorganisms 11, no. 2: 349. https://doi.org/10.3390/microorganisms11020349

APA Style

Xie, L., Yoshida, N., & Meng, L. (2023). Polyphasic Characterization of Geotalea uranireducens NIT-SL11 Newly Isolated from a Complex of Sewage Sludge and Microbially Reduced Graphene Oxide. Microorganisms, 11(2), 349. https://doi.org/10.3390/microorganisms11020349

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop