scholarly journals Bacterial Microcompartment-Dependent 1,2-Propanediol Utilization Stimulates Anaerobic Growth of Listeria monocytogenes EGDe

2019 ◽  
Vol 10 ◽  
Author(s):  
Zhe Zeng ◽  
Eddy J. Smid ◽  
Sjef Boeren ◽  
Richard A. Notebaart ◽  
Tjakko Abee
Keyword(s):  
Listeria Monocytogenes ◽  
Anaerobic Growth ◽  
Bacterial Microcompartment

scholarly journals Anaerobic Growth of Listeria monocytogenes on Rhamnose Is Stimulated by Vitamin B 12 and Bacterial Microcompartment-Dependent 1,2-Propanediol Utilization

mSphere ◽  
2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Molecular Mechanisms ◽  
Foodborne Pathogen ◽  
Anaerobic Growth ◽  
Severe Illness ◽  
Vitamin B ◽  
Anaerobic Conditions ◽  
Content Type ◽  
Bacterial Microcompartment ◽  
Vitamin B 12

Listeria monocytogenes is a foodborne pathogen causing severe illness and, as such, it is crucial to understand the molecular mechanisms contributing to its survival strategy and pathogenicity. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol.

scholarly journals Impact of vitamin B12 on rhamnose metabolism, stress defense and in-vitro virulence of Listeria monocytogenes

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Growth Stimulation ◽  
Anaerobic Growth ◽  
Salmonella Spp ◽  
Defense Proteins ◽  
Bacterial Microcompartment ◽  
The Impact ◽  
Stress Defense

Listeria monocytogenes is a facultative anaerobe which can cause a severe food-borne infection known as listeriosis. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded by L. monocytogenes in aerobic and anaerobic conditions into lactate, acetate and 1,2-propanediol. Our previous study showed that addition of vitamin B12 stimulates anaerobic growth of L. monocytogenes on rhamnose due to the activation of bacterial microcompartment (BMC)-dependent 1,2-propanediol utilization with concomitant production of propionate and propanol. Notably, anaerobic propanediol metabolism has been linked to virulence of enteric pathogens including Salmonella spp. and L. monocytogenes. In this study we investigate the impact of B12 on aerobic and anerobic growth of L. monocytogenes on rhamnose, and observed growth stimulation and pdu BMC activation only in anaerobically grown cells with B12 added to the medium. Comparative Caco-2 virulence assays, showed that these pdu BMC induced cells have significantly higher translocation efficiency compared to aerobically grown cells (without and with added B12) and non-induced anaerobically grown cells, while adhesion and invasion capacity is similar for all cells. Comparative proteomics analysis showed specific and overlapping responses linked to metabolic shifts, activation of stress defense proteins and virulence factors, with RNA polymerase sigma factor SigL; teichoic acids export ATP-binding protein, TagH; DNA repair and protection proteins RadA and DPS; and glutathione synthase GshAB previously linked to activation of virulence response in L. monocytogenes, uniquely upregulated in anaerobically rhamnose grown pdu BMC induced cells. Our results shed new light into B12 impact on L. monocytogenes competitive fitness and virulence.

scholarly journals Anaerobic growth of Listeria monocytogenes on rhamnose is stimulated by Vitamin B12 and bacterial microcompartment dependent 1,2-propanediol utilization

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Anaerobic Growth ◽  
Physiological Effects ◽  
Human Intestine ◽  
Human Pathogens ◽  
Bacterial Microcompartment ◽  
Food Borne ◽  
Transmission Electron ◽  
Shell Proteins

AbstractThe food-borne pathogen Listeria monocytogenes is able to form proteinaceous organelles called bacterial microcompartments (BMCs) that optimize the utilization of substrates, such as 1,2-propanediol, and confer an anaerobic growth advantage. Rhamnose is a deoxyhexose sugar abundant in a range of environments including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol, next to acetate and lactate. Rhamnose-derived 1,2-propanediol has been found to link with BMCs in a limited number of commensal human colonic species and some human pathogens such as Salmonella enterica, but the involvement of BMCs in rhamnose metabolism and potential physiological effects on L. monocytogenes are still unknown. In this study, we firstly test the effect of rhamnose uptake and utilization on anaerobic growth of L. monocytogenes EGDe without and with added vitamin B12, followed by metabolic analysis. We unveil that the vitamin B12-dependent activation of pdu stimulates metabolism and anaerobic growth of L. monocytogenes EGDe on rhamnose via 1,2-propanediol degradation into 1-propanol and propionate. Transmission electron microscopy of pdu-induced cells shows that BMCs are formed and additional proteomics experiments confirm expression of pdu BMC shell proteins and enzymes. Finally, we discuss physiological effects and energy efficiency of L. monocytogenes pdu BMC-driven anaerobic rhamnose metabolism and impact on competitive fitness in environments such as the human intestine.

scholarly journals Impact of bacterial microcompartment-dependent ethanolamine and propanediol metabolism on Listeria monocytogenes interactions with Caco-2 cells

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Human Cells ◽  
Cellular Systems ◽  
Anaerobic Growth ◽  
Commensal Microbiota ◽  
Bacterial Microcompartment ◽  
Transmission Electron ◽  
Well Assay ◽  
Phospholipid Degradation

AbstractBacterial microcompartment (BMC) dependent ethanolamine (eut) and propanediol utilization (pdu) has recently been shown to stimulate anaerobic growth of Listeria monocytogenes. This metabolic repertoire conceivably contributes to the competitive fitness of L. monocytogenes in the human gastrointestinal (GI) tract, where these compounds become available following phospholipid degradation and mucus-derived rhamnose metabolism by commensal microbiota. Previous transcriptomics and mutant studies of eut and pdu L. monocytogenes suggested a possible role of eut and pdu BMC metabolism in transmission in foods and pathogenicity, but data on a potential role of L. monocytogenes interaction with human cells is currently absent. First, we ask which cellular systems are expressed in the activation of eut and pdu BMC metabolism and the extent to which these systems are conserved between the states. We find common and unique systems related to metabolic shifts, stress and virulence factors. Next, we hypothesize that these common and unique activated cellular systems contribute to a role in the interaction of L. monocytogenes interaction with human cells. We present evidence that metabolically primed L. monocytogenes with active eut and pdu BMCs, as confirmed by metabolic analysis, transmission electron microscopy and proteomics, show significantly enhanced translocation efficacy compared to non-induced cells in a trans-well assay using Caco-2 cells, while adhesion and invasion capacity was similar. Taken together, our results provide insights into the possible key cellular players that drive translocation efficacy upon eut and pdu BMC activation.

scholarly journals The Nonmevalonate Pathway of Isoprenoid Biosynthesis Supports Anaerobic Growth of Listeria monocytogenes

2019 ◽  
Vol 88 (2) ◽  
Author(s):  
Eric D. Lee ◽  
Kathleen I. Navas ◽  
Daniel A. Portnoy
Keyword(s):  
Growth Rate ◽  
Listeria Monocytogenes ◽  
Doubling Time ◽  
Mevalonate Pathway ◽  
Anaerobic Growth ◽  
Clinical Isolates ◽  
Content Type ◽  
Nonmevalonate Pathway ◽  
Common Precursor ◽  
The Difference

ABSTRACT Isoprenoids are an essential and diverse class of molecules, present in all forms of life, that are synthesized from an essential common precursor derived from either the mevalonate pathway or the nonmevalonate pathway. Most bacteria have one pathway or the other, but the Gram-positive, facultative intracellular pathogen Listeria monocytogenes is unusual because it carries all the genes for both pathways. While the mevalonate pathway has previously been reported to be essential, here we demonstrate that the nonmevalonate pathway can support growth of strains 10403S and EGD-e, but only anaerobically. L. monocytogenes lacking the gene hmgR, encoding the rate-limiting enzyme of the mevalonate pathway, had a doubling time of 4 h under anaerobic conditions, in contrast to the 45 min doubling time of the wild type. In contrast, deleting hmgR in two clinical isolates resulted in mutants that grew significantly faster, doubling in approximately 2 h anaerobically, although they still failed to grow under aerobic conditions without mevalonate. The difference in anaerobic growth rate was traced to three amino acid changes in the nonmevalonate pathway enzyme GcpE, and these changes were sufficient to increase the growth rate of 10403S to the rate observed in the clinical isolates. Despite an increased growth rate, virulence was still dependent on the mevalonate pathway in 10403S strains expressing the more active GcpE allele.

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 Identification of genes essential for anaerobic growth of Listeria monocytogenes

Microbiology ◽  
2014 ◽  
Vol 160 (4) ◽  
pp. 752-765 ◽  
Author(s):  
Stefanie Müller-Herbst ◽  
Stefanie Wüstner ◽  
Anna Mühlig ◽  
Daniela Eder ◽  
Thilo M. Fuchs ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Oxygen Tension ◽  
Dna Microarrays ◽  
Growth Conditions ◽  
Anaerobic Growth ◽  
Phenotypic Analysis ◽  
Knock Out ◽  
Metabolic Genes ◽  
Lower Oxygen ◽  
Bacterial Fitness

The facultative anaerobic bacterium Listeria monocytogenes encounters microaerophilic or anaerobic conditions in various environments, e.g. in soil, in decaying plant material, in food products and in the host gut. To elucidate the adaptation of Listeria monocytogenes to variations in oxygen tension, global transcription analyses using DNA microarrays were performed. In total, 139 genes were found to be transcribed differently during aerobic and anaerobic growth; 111 genes were downregulated and 28 genes were upregulated anaerobically. The oxygen-dependent transcription of central metabolic genes is in agreement with results from earlier physiological studies. Of those genes more strongly expressed under lower oxygen tension, 20 were knocked out individually. Growth analysis of these knock out mutants did not indicate an essential function for the respective genes during anaerobiosis. However, even if not essential, transcriptional induction of several genes might optimize the bacterial fitness of Listeria monocytogenes in anaerobic niches, e.g. during colonization of the gut. For example, expression of the anaerobically upregulated gene lmo0355, encoding a fumarate reductase α chain, supported growth on 10 mM fumarate under anaerobic but not under aerobic growth conditions. Genes essential for anaerobic growth were identified by screening a mutant library. Eleven out of 1360 investigated mutants were sensitive to anaerobiosis. All 11 mutants were interrupted in the atp locus. These results were further confirmed by phenotypic analysis of respective in-frame deletion and complementation mutants, suggesting that the generation of a proton motive force via F1F0-ATPase is essential for anaerobic proliferation of Listeria monocytogenes.

Heat Stability of Virulence-Associated Enzymes from Listeria monocytogenes SLCC 5764

1998 ◽  
Vol 61 (7) ◽  
pp. 899-902 ◽  
Author(s):  
RACHEL B. ZEMSER ◽  
SCOTT E. MARTIN
Keyword(s):  
Listeria Monocytogenes ◽  
Elevated Temperatures ◽  
Heat Stability ◽  
Anaerobic Growth ◽  
Listeriolysin O ◽  
Optimum Growth ◽  
Anaerobic Conditions ◽  
Optimum Growth Temperature ◽  
Heat Stable ◽  
Almost All

The enzyme activities of listeriolysin O (LLO), phosphatidylinositol-specific phospholipase C (PI-PLC), catalase (CA), and superoxide dismutase (SOD) from Listeria monocytogenes SLCC 5764 were examined after heat treatment, growth at elevated temperatures, and anaerobic growth. Growth at elevated temperatures may influence virulence as expressed by virulence enzyme activity. The enzymes were heated for 0 and 10 min at temperatures ranging from 40 to 100°C. The production of LLO was examined when the bacterium was grown at elevated temperatures, and the production of LLO, CA, and SOD were examined after anaerobic growth. LLO was the most heat-labile factor, losing almost all activity when heated above 50°C. CA was more heat stable, showing no decrease in activity until it was heated at 55°C. The PI-PLC enzyme was most heat resistant: the activity decreased between 40 and 50°C and continued to decrease when heated between 65 and 100°C. When L. monocytogenes was grown at elevated temperatures, it produced less LLO than when grown at the optimum growth temperature of 37°C. When the organism was grown under anaerobic conditions, the levels of CA and LLO decreased, while the SOD level remained unchanged.

Survival of Staphylococcus aureus and Listeria monocytogenes on Vacuum-Packaged Beef Jerky and Related Products Stored at 21°C

2006 ◽  
Vol 69 (9) ◽  
pp. 2263-2267 ◽  
Author(s):  
STEVEN C. INGHAM ◽  
GINA SEARLS ◽  
SUNISH MOHANAN ◽  
DENNIS R. BUEGE
Keyword(s):  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Pathogenic Bacteria ◽  
Room Temperature ◽  
Anaerobic Growth ◽  
Department Of Agriculture ◽  
Small Individual ◽  
Beef Jerky ◽  
Maximum Water ◽  
Packaged Beef

In the manufacture of beef jerky, a thermal lethality step is followed by drying to prevent growth of pathogenic bacterial postprocessing contaminants on the finished product. Recent guidelines from the U.S. Department of Agriculture have raised the question of the maximum water activity (aw) in jerky products that will inhibit growth of pathogenic bacteria. The survival of the potential postprocessing contaminants Staphylococcus aureus and Listeria monocytogenes was evaluated on 15 vacuum-packaged beef jerky and related products with aw values ranging from 0.47 to 0.87, just below the 0.88 limit reported for anaerobic growth of S. aureus. Small individual product pieces were inoculated on the outer surface with five strains each of S. aureus and L. monocytogenes, repackaged under vacuum, and stored at room temperature (21°C) for 4 weeks. Pathogen numbers were determined before storage and after 1 and 4 weeks. None of the 15 jerky products supported growth of either pathogen. Counts of S. aureus fell by 0.2 to 1.8 log CFU after 1 week of storage and by 0.6 to 5.3 log CFU after 4 weeks of storage. Numbers of L. monocytogenes fell by 0.6 to 4.7 log CFU and by 2.3 to 5.6 log CFU after 1 and 4 weeks of storage, respectively. Although factors other than aw may have some effect on pathogen survival, the results of the present study clearly support drying beef jerky to an aw of ≤0.87 to ensure that bacterial pathogens cannot grow on vacuum-packaged product stored at room temperature.

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