scholarly journals Bacterial microcompartments linked to the flavin-based extracellular electron transfer drives anaerobic ethanolamine utilization in Listeria monocytogenes

2020 ◽  
Author(s):  
Zhe Zeng ◽  
Sjef Boeren ◽  
Varaang Bhandula ◽  
Samuel H. Light ◽  
Eddy J. Smid ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Listeria Monocytogenes ◽  
Electron Acceptor ◽  
Cell Membranes ◽  
Anaerobic Growth ◽  
Extracellular Electron Transfer ◽  
Severe Illness ◽  
Redox Imbalance ◽  
Bacterial Microcompartments

AbstractEthanolamine (EA) is a valuable microbial carbon and nitrogen source derived from phospholipids present in cell membranes. EA catabolism is suggested to occur in so-called bacterial microcompartments (BMCs) and activation of EA utilization (eut) genes is linked to bacterial pathogenesis. Despite reports showing that activation of eut in Listeria monocytogenes is regulated by a vitamin B12-binding riboswitch and that upregulation of eut genes occurs in mice, it remains unknown whether EA catabolism is BMC dependent. Here, we provide evidence for BMC-dependent anaerobic EA utilization via metabolic analysis, proteomics and electron microscopy. First, we show B12-induced activation of the eut operon in L. monocytogenes coupled to uptake and utilization of EA thereby enabling growth. Next, we demonstrate BMC formation in conjunction to EA catabolism with the production of acetate and ethanol in a molar ratio of 2:1. Flux via the ATP generating acetate branch causes an apparent redox imbalance due to reduced regeneration of NAD+ in the ethanol branch resulting in a surplus of NADH. We hypothesize that the redox imbalance is compensated by linking eut BMC to anaerobic flavin-based extracellular electron transfer (EET). Using L. monocytogenes wild type, a BMC mutant and a EET mutant, we demonstrate an interaction between BMC and EET and provide evidence for a role of Fe3+ as an electron acceptor. Taken together, our results suggest an important role of anaerobic BMC-dependent EA catabolism in the physiology of L. monocytogenes, with a crucial role for the flavin-based EET system in redox balancing.IMPORTANCEListeria monocytogenes is a food-borne pathogen causing severe illness and, as such, it is crucial to understand the molecular mechanisms contributing to pathogenicity. One carbon source that allows L. monocytogenes to grow in humans is ethanolamine (EA), which is derived from phospholipids present in eukaryotic cell membranes. It is hypothesized that EA utilization occurs in bacterial microcompartments (BMCs), self-assembling subcellular proteinaceous structures and analogs of eukaryotic organelles. Here, we demonstrate that BMC-driven utilization of EA in L. monocytogenes results in increased energy production essential for anaerobic growth. However, exploiting BMCs and the encapsulated metabolic pathways also requires balancing of oxidative and reductive pathways. We now provide evidence that L. monocytogenes copes with this by linking BMC activity to flavin-based extracellular electron transfer (EET) using iron as an electron acceptor. Our results shed new light on an important molecular mechanism that enables L. monocytogenes to grow using host-derived phospholipid degradation products.

scholarly journals Bacterial Microcompartments Coupled with Extracellular Electron Transfer Drive the Anaerobic Utilization of Ethanolamine in Listeria monocytogenes

mSystems ◽  
2021 ◽  
Vol 6 (2) ◽  
Author(s):  
Zhe Zeng ◽  
Sjef Boeren ◽  
Varaang Bhandula ◽  
Samuel H. Light ◽  
Eddy J. Smid ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Listeria Monocytogenes ◽  
Electron Acceptor ◽  
Cell Membranes ◽  
Extracellular Electron Transfer ◽  
Vitamin B ◽  
Content Type ◽  
Redox Imbalance ◽  
Bacterial Microcompartments

ABSTRACT Ethanolamine (EA) is a valuable microbial carbon and nitrogen source derived from cell membranes. EA catabolism is suggested to occur in a cellular metabolic subsystem called a bacterial microcompartment (BMC), and the activation of EA utilization (eut) genes is linked to bacterial pathogenesis. Despite reports showing that the activation of eut is regulated by a vitamin B12-binding riboswitch and that upregulation of eut genes occurs in mice, it remains unknown whether EA catabolism is BMC dependent in Listeria monocytogenes. Here, we provide evidence for BMC-dependent anaerobic EA utilization via metabolic analysis, proteomics, and electron microscopy. First, we show vitamin B12-induced activation of the eut operon in L. monocytogenes coupled to the utilization of EA, thereby enabling growth. Next, we demonstrate BMC formation connected with EA catabolism with the production of acetate and ethanol in a molar ratio of 2:1. Flux via the ATP-generating acetate branch causes an apparent redox imbalance due to the reduced regeneration of NAD+ in the ethanol branch resulting in a surplus of NADH. We hypothesize that the redox imbalance is compensated by linking eut BMCs to anaerobic flavin-based extracellular electron transfer (EET). Using L. monocytogenes wild-type, BMC mutant, and EET mutant strains, we demonstrate an interaction between BMCs and EET and provide evidence for a role of Fe3+ as an electron acceptor. Taken together, our results suggest an important role of BMC-dependent EA catabolism in L. monocytogenes growth in anaerobic environments like the human gastrointestinal tract, with a crucial role for the flavin-based EET system in redox balancing. IMPORTANCE Listeria monocytogenes is a foodborne pathogen causing severe illness, and as such, it is crucial to understand the molecular mechanisms contributing to pathogenicity. One carbon source that allows L. monocytogenes to grow in humans is ethanolamine (EA), which is derived from phospholipids present in eukaryotic cell membranes. It is hypothesized that EA utilization occurs in bacterial microcompartments (BMCs), self-assembling subcellular proteinaceous structures and analogs of eukaryotic organelles. Here, we demonstrate that BMC-driven utilization of EA in L. monocytogenes results in increased energy production essential for anaerobic growth. However, exploiting BMCs and the encapsulated metabolic pathways also requires the balancing of oxidative and reductive pathways. We now provide evidence that L. monocytogenes copes with this by linking BMC activity to flavin-based extracellular electron transfer (EET) using iron as an electron acceptor. Our results shed new light on an important molecular mechanism that enables L. monocytogenes to grow using host-derived phospholipid degradation products.

Impact of Graphitic Structures in Pyrogenic Carbon on Microbial Reduction of Ferrihydrite

2021 ◽  
Author(s):  
wentao yu ◽  
baoliang chen
Keyword(s):  
Electron Transfer ◽  
Pyrolysis Temperature ◽  
Shewanella Oneidensis ◽  
Exchange Capacity ◽  
Microbial Reduction ◽  
Systematic Evaluation ◽  
Extracellular Electron Transfer ◽  
Pyrogenic Carbon ◽  
The Impact

<p>Pyrogenic carbon plays important roles in microbial reduction of ferrihydrite by shuttling electrons in the extracellular electron transfer (EET) processes. Despite its importance, a full assessment on the impact of graphitic structures in pyrogenic carbon on microbial reduction of ferrihydrite has not been conducted. This study is a systematic evaluation of microbial ferrihydrite reduction by Shewanella oneidensis MR-1 in the presence of pyrogenic carbon with various graphitization extents. The results showed that the rates and extents of microbial ferrihydrite reduction were significantly enhanced in the presence of pyrogenic carbon, and increased with increasing pyrolysis temperature. Combined spectroscopic and electrochemical analyses suggested that the rate of microbial ferrihydrite reduction were dependent on the electrical conductivity of pyrogenic carbon (i.e., graphitization extent), rather than the electron exchange capacity. The key role of graphitic structures in pyrogenic carbon in mediating EET was further evidenced by larger microbial electrolysis current with pyrogenic carbon prepared at higher pyrolysis temperatures. This study provides new insights into the electron transfer in the pyrogenic carbon-mediated microbial reduction of ferrihydrite.</p>

Wood derived biochar as electron donor and its influence on microbial denitrification: Role of extracellular polymeric substances in extracellular electron transfer

2020 ◽  
Author(s):  
Kuppusamy Sathishkumar ◽  
Yi Li ◽  
Rana Muhammad Adnan Ikram
Keyword(s):  
Electron Transfer ◽  
Electron Donor ◽  
Electrochemical Behavior ◽  
Nitrate Reduction ◽  
Extracellular Polymeric Substances ◽  
Electrochemical Impedance ◽  
Electrochemical Analysis ◽  
Extracellular Electron Transfer ◽  
Microbial Denitrification

<p>Biochar is extensively used in environmental pollutant remediation because of its diverse property, however the effect of biochar on microbial nitrate reduction and electrochemical behavior of biochar remain unknown. Also electron transfer from the microbial cells to electron donor or acceptor have been transport across the extracellular polymeric substances (EPS), however it was unclear whether extracellular polymeric substances captured or enhance the electrons.  Hence, aim of the present study is to investigate the electrochemical behavior of biochar and its effects on microbial nitrate reduction and elucidate the role of extracellular polymeric substances in extracellular electron transfer (EET).  The biochar was prepared at different pyrolysis temperatures (400 °C, 500 °C and 600 °C) and their electrochemical behavior was characterized by electrochemical analysis (cyclic voltammetry, electrochemical impedance spectrum, chronoamperometry). Results demonstrated that all the biochars could donate and accept the electrons, impact of biochar on microbial nitrate reduction was studied and the results showed that biochar prepared at 400 °C significantly enhances microbial nitrate reduction process. Phenol O-H and quinone C=O surface functional groups on the biochar contributes in the overall electron exchange which accelerated the nitrate reduction. The role of EPS in EET by electrochemical analysis results reveals that outer membrane c-type cytochrome and flavin protein from the biofilm was involved in electron transfer process, and EPS act as transient media for microbial EET. Overall, present study suggested that biochar could be used as eco-friendly material for the enhancement of microbial denitrification.</p>

scholarly journals Geothrix fermentans Secretes Two Different Redox-Active Compounds To Utilize Electron Acceptors across a Wide Range of Redox Potentials

2012 ◽  
Vol 78 (19) ◽  
pp. 6987-6995 ◽  
Author(s):  
Misha G. Mehta-Kolte ◽  
Daniel R. Bond
Keyword(s):  
Electron Transfer ◽  
Redox Potential ◽  
Electron Acceptor ◽  
Extracellular Electron Transfer ◽  
Redox Potentials ◽  
Active Compounds ◽  
Content Type ◽  
Electron Shuttles ◽  
Redox Active ◽  
Geothrix Fermentans

ABSTRACTThe current understanding of dissimilatory metal reduction is based primarily on isolates from the proteobacterial generaGeobacterandShewanella. However, environments undergoing active Fe(III) reduction often harbor less-well-studied phyla that are equally abundant. In this work, electrochemical techniques were used to analyze respiratory electron transfer by the only known Fe(III)-reducing representative of theAcidobacteria,Geothrix fermentans. In contrast to previously characterized metal-reducing bacteria, which typically reach maximal rates of respiration at electron acceptor potentials of 0 V versus standard hydrogen electrode (SHE),G. fermentansrequired potentials as high as 0.55 V to respire at its maximum rate. In addition,G. fermentanssecreted two different soluble redox-active electron shuttles with separate redox potentials (−0.2 V and 0.3 V). The compound with the lower midpoint potential, responsible for 20 to 30% of electron transfer activity, was riboflavin. The behavior of the higher-potential compound was consistent with hydrophilic UV-fluorescent molecules previously found inG. fermentanssupernatants. Both electron shuttles were also produced when cultures were grown with Fe(III), but not when fumarate was the electron acceptor. This study reveals thatGeothrixis able to take advantage of higher-redox-potential environments, demonstrates that secretion of flavin-based shuttles is not confined toShewanella, and points to the existence of high-potential-redox-active compounds involved in extracellular electron transfer. Based on differences between the respiratory strategies ofGeothrixandGeobacter, these two groups of bacteria could exist in distinctive environmental niches defined by redox potential.

scholarly journals Extracellular electron transfer-dependent anaerobic oxidation of ammonium by anammox bacteria

2019 ◽  
Author(s):  
Dario R. Shaw ◽  
Muhammad Ali ◽  
Krishna P. Katuri ◽  
Jeffrey A. Gralnick ◽  
Joachim Reimann ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Electron Acceptor ◽  
Electron Acceptors ◽  
Extracellular Electron Transfer ◽  
Anammox Bacteria ◽  
Significant Finding ◽  
Ammonium Oxidation ◽  
Anaerobic Oxidation ◽  
Terminal Electron Acceptor ◽  
Carbon Based

AbstractAnaerobic ammonium oxidation (anammox) by anammox bacteria contributes significantly to the global nitrogen cycle, and plays a major role in sustainable wastewater treatment. Anammox bacteria convert ammonium (NH4+) to dinitrogen gas (N2) using nitrite (NO2−) or nitric oxide (NO) as the electron acceptor. In the absence of NO2− or NO, anammox bacteria can couple formate oxidation to the reduction of metal oxides such as Fe(III) or Mn(IV). Their genomes contain homologs of Geobacter and Shewanella cytochromes involved in extracellular electron transfer (EET). However, it is still unknown whether anammox bacteria have EET capability and can couple the oxidation of NH4+ with transfer of electrons to carbon-based insoluble extracellular electron acceptors. Here we show using complementary approaches that in the absence of NO2−, freshwater and marine anammox bacteria couple the oxidation of NH4+ with transfer of electrons to carbon-based insoluble extracellular electron acceptors such as graphene oxide (GO) or electrodes poised at a certain potential in microbial electrolysis cells (MECs). Metagenomics, fluorescence in-situ hybridization and electrochemical analyses coupled with MEC performance confirmed that anammox electrode biofilms were responsible for current generation through EET-dependent oxidation of NH4+. 15N-labelling experiments revealed the molecular mechanism of the EET-dependent anammox process. NH4+ was oxidized to N2 via hydroxylamine (NH2OH) as intermediate when electrode was the terminal electron acceptor. Comparative transcriptomics analysis supported isotope labelling experiments and revealed an alternative pathway for NH4+ oxidation coupled to EET when electrode is used as electron acceptor compared to NO2−as electron acceptor. To our knowledge, our results provide the first experimental evidence that marine and freshwater anammox bacteria can couple NH4+ oxidation with EET, which is a significant finding, and challenges our perception of a key player of anaerobic oxidation of NH4+ in natural environments and engineered systems.

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