scholarly journals Chaperones mainly suppress primary nucleation during formation of functional amyloid required for bacterial biofilm formation

2022 ◽  
Author(s):  
Madhu Nagaraj ◽  
Zahra Najarzadeh ◽  
Jonathan Pansieri ◽  
Ludmilla A. Morozova-Roche ◽  
Henrik Biverstål ◽  
...  
Keyword(s):  
Neurodegenerative Diseases ◽  
Biofilm Formation ◽  
Bacterial Biofilm ◽  
Functional Amyloid ◽  
E Coli ◽  
Primary Nucleation ◽  
Functional Amyloids

Unlike misfolding in neurodegenerative diseases, aggregation of functional amyloids involved in bacterial biofilm, e.g. CsgA (E. coli) and FapC (Pseudomonas), is carefully regulated. However, it is unclear whether functional aggregation...

scholarly journals More than Enzymes That Make or Break Cyclic Di-GMP—Local Signaling in the Interactome of GGDEF/EAL Domain Proteins of Escherichia coli

mBio ◽  
2017 ◽  
Vol 8 (5) ◽  
Author(s):  
Olga Sarenko ◽  
Gisela Klauck ◽  
Franziska M. Wilke ◽  
Vanessa Pfiffer ◽  
Anja M. Richter ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Second Messenger ◽  
Bacterial Biofilm ◽  
Interaction Patterns ◽  
Systematic Analysis ◽  
E Coli ◽  
Diguanylate Cyclases ◽  
Hub Proteins ◽  
Effector Target

ABSTRACT The bacterial second messenger bis-(3′-5′)-cyclic diguanosine monophosphate (c-di-GMP) ubiquitously promotes bacterial biofilm formation. Intracellular pools of c-di-GMP seem to be dynamically negotiated by diguanylate cyclases (DGCs, with GGDEF domains) and specific phosphodiesterases (PDEs, with EAL or HD-GYP domains). Most bacterial species possess multiple DGCs and PDEs, often with surprisingly distinct and specific output functions. One explanation for such specificity is “local” c-di-GMP signaling, which is believed to involve direct interactions between specific DGC/PDE pairs and c-di-GMP-binding effector/target systems. Here we present a systematic analysis of direct protein interactions among all 29 GGDEF/EAL domain proteins of Escherichia coli . Since the effects of interactions depend on coexpression and stoichiometries, cellular levels of all GGDEF/EAL domain proteins were also quantified and found to vary dynamically along the growth cycle. Instead of detecting specific pairs of interacting DGCs and PDEs, we discovered a tightly interconnected protein network of a specific subset or “supermodule” of DGCs and PDEs with a coregulated core of five hyperconnected hub proteins. These include the DGC/PDE proteins representing the c-di-GMP switch that turns on biofilm matrix production in E. coli . Mutants lacking these core hub proteins show drastic biofilm-related phenotypes but no changes in cellular c-di-GMP levels. Overall, our results provide the basis for a novel model of local c-di-GMP signaling in which a single strongly expressed master PDE, PdeH, dynamically eradicates global effects of several DGCs by strongly draining the global c-di-GMP pool and thereby restricting these DGCs to serving as local c-di-GMP sources that activate specific colocalized effector/target systems. IMPORTANCE c-di-GMP signaling in bacteria is believed to occur via changes in cellular c-di-GMP levels controlled by antagonistic and potentially interacting pairs of diguanylate cyclases (DGCs) and c-di-GMP phosphodiesterases (PDEs). Our systematic analysis of protein-protein interaction patterns of all 29 GGDEF/EAL domain proteins of E. coli , together with our measurements of cellular c-di-GMP levels, challenges both aspects of this current concept. Knocking out distinct DGCs and PDEs has drastic effects on E. coli biofilm formation without changing the cellular c-di-GMP level. In addition, rather than generally coming in interacting DGC/PDE pairs, a subset of DGCs and PDEs operates as central interaction hubs in a larger "supermodule," with other DGCs and PDEs behaving as “lonely players” without contacts to other c-di-GMP-related enzymes. On the basis of these data, we propose a novel concept of “local” c-di-GMP signaling in bacteria with multiple enzymes that make or break the second messenger c-di-GMP.

scholarly journals RNase I regulates Escherichia coli 2′,3′-cyclic nucleotide monophosphate levels and biofilm formation

2018 ◽  
Vol 475 (8) ◽  
pp. 1491-1506 ◽  
Author(s):  
Benjamin M. Fontaine ◽  
Kevin S. Martin ◽  
Jennifer M. Garcia-Rodriguez ◽  
Claire Jung ◽  
Laura Briggs ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Dna Replication ◽  
Cyclic Nucleotide ◽  
Biofilm Formation ◽  
Physiological Role ◽  
Rna Degradation ◽  
Bacterial Biofilm ◽  
Biological Functions ◽  
Salvage Pathway ◽  
E Coli

Regulation of nucleotide and nucleoside concentrations is critical for faithful DNA replication, transcription, and translation in all organisms, and has been linked to bacterial biofilm formation. Unusual 2′,3′-cyclic nucleotide monophosphates (2′,3′-cNMPs) recently were quantified in mammalian systems, and previous reports have linked these nucleotides to cellular stress and damage in eukaryotes, suggesting an intriguing connection with nucleotide/nucleoside pools and/or cyclic nucleotide signaling. This work reports the first quantification of 2′,3′-cNMPs in Escherichia coli and demonstrates that 2′,3′-cNMP levels in E. coli are generated specifically from RNase I-catalyzed RNA degradation, presumably as part of a previously unidentified nucleotide salvage pathway. Furthermore, RNase I and 2′,3′-cNMP levels are demonstrated to play an important role in controlling biofilm formation. This work identifies a physiological role for cytoplasmic RNase I and constitutes the first progress toward elucidating the biological functions of bacterial 2′,3′-cNMPs.

scholarly journals Disrupting Irreversible Bacterial Adhesion and Biofilm Formation with an Engineered Enzyme

2021 ◽  
Author(s):  
Holly M. Mayton ◽  
Sharon L. Walker ◽  
Bryan W. Berger
Keyword(s):  
Listeria Monocytogenes ◽  
Bacterial Adhesion ◽  
Biofilm Formation ◽  
Glycosyl Hydrolase ◽  
Cell Surface Hydrophobicity ◽  
Bacterial Biofilm ◽  
Enzyme Treatment ◽  
E Coli ◽  
Initial Adhesion ◽  
The Impact

Biofilm formation is often attributed to post-harvest bacteria persistence on fresh produce and food handling surfaces. In this study, a predicted glycosyl hydrolase enzyme was expressed, purified and validated for removal of microbial biofilms from biotic and abiotic surfaces under conditions used for chemical cleaning agents. Crystal violet biofilm staining assays revealed that 0.1 mg/mL of enzyme inhibited up to 41% of biofilm formation by E. coli O157:H7, E. coli 25922, Salmonella Typhimurium, and Listeria monocytogenes. Further, the enzyme was effective at removing mature biofilms, providing a 35% improvement over rinsing with a saline solution alone. Additionally, a parallel-plate flow cell was used to directly observe and quantify the impact of enzyme rinses on E. coli O157:H7 cells adhered to spinach leaf surfaces. The presence of 1 mg/L enzyme resulted in nearly 6 times greater detachment rate coefficients than a DI water rinse, while the total cells removed from the surface increased from 10% to 25% over the 30 minute rinse time, reversing the initial phases of biofilm formation. Enzyme treatment of all 4 cell types resulted in significantly reduced cell surface hydrophobicity, and collapse of negatively stained E. coli 25922 cells imaged by electron microscopy, suggesting potential polysaccharide surface modification of enzyme-treated bacteria. Collectively, these results point to the broad substrate specificity and robustness of the enzyme to different types of biofilm stages, solution conditions and pathogen biofilm types, and may be useful as a method for removal or inhibition of bacterial biofilm formation. IMPORTANCE In this study, the ability of an engineered enzyme to reduce bacterial adhesion and biofilm formation of several foodborne pathogens was demonstrated, representing a promising option for enhancing or replacing chlorine and other chemical sanitizers in food processing applications. Specifically, significant reductions of the pathogens Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms are observed, as well as reduction in initial adhesion. Enzymes have the added benefits of being green, sustainable alternatives to chemical sanitizers, as well as having minimal impact on food properties, in contrast with many alternative antimicrobial options such as bleach that aim to minimize food safety risks.

scholarly journals Cross-talk between individual phenol-soluble modulins in Staphylococcus aureus biofilm enables rapid and efficient amyloid formation

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Masihuz Zaman ◽  
Maria Andreasen
Keyword(s):  
Staphylococcus Aureus ◽  
Self Assembly ◽  
Biofilm Formation ◽  
Opportunistic Pathogen ◽  
Amyloid Formation ◽  
Secondary Nucleation ◽  
Primary Nucleation ◽  
Functional Amyloids ◽  
High Degree

The infective ability of the opportunistic pathogen Staphylococcus aureus, recognized as the most frequent cause of biofilm-associated infections, is associated with biofilm-mediated resistance to host immune response. Phenol-soluble modulins (PSM) comprise the structural scaffold of S. aureus biofilms through self-assembly into functional amyloids, but the role of individual PSMs during biofilm formation remains poorly understood and the molecular pathways of PSM self-assembly are yet to be identified. Here we demonstrate high degree of cooperation between individual PSMs during functional amyloid formation. PSMα3 initiates the aggregation, forming unstable aggregates capable of seeding other PSMs resulting in stable amyloid structures. Using chemical kinetics we dissect the molecular mechanism of aggregation of individual PSMs showing that PSMα1, PSMα3 and PSMβ1 display secondary nucleation whereas PSMβ2 aggregates through primary nucleation and elongation. Our findings suggest that various PSMs have evolved to ensure fast and efficient biofilm formation through cooperation between individual peptides.

scholarly journals Antimicrobial Activity and Prevention of Bacterial Biofilm Formation of Silver and Zinc Oxide Nanoparticle-Containing Polyester Surfaces at Various Concentrations for Use

Foods ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 442 ◽  
Author(s):  
Fabio Fontecha-Umaña ◽  
Abel Guillermo Ríos-Castillo ◽  
Carolina Ripolles-Avila ◽  
José Juan Rodríguez-Jerez
Keyword(s):  
Antimicrobial Activity ◽  
Zinc Oxide ◽  
Biofilm Formation ◽  
Antimicrobial Properties ◽  
Bacterial Biofilm ◽  
Antimicrobial Efficacy ◽  
Ag Nps ◽  
Zno Nps ◽  
E Coli ◽  
Food Contact Surfaces

Food contact surfaces are primary sources of bacterial contamination in food industry processes. With the objective of preventing bacterial adhesion and biofilm formation on surfaces, this study evaluated the antimicrobial activity of silver (Ag-NPs) and zinc oxide (ZnO-NPs) nanoparticle-containing polyester surfaces (concentration range from 400 ppm to 850 ppm) using two kinds of bacteria, Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli), and the prevention of bacterial biofilm formation using the pathogen Listeria monocytogenes. The results of antimicrobial efficacy (reductions ≥ 2 log CFU/cm2) showed that at a concentration of 850 ppm, ZnO-NPs were effective against only E. coli (2.07 log CFU/cm2). However, a concentration of 400 ppm of Ag-NPs was effective against E. coli (4.90 log CFU/cm2) and S. aureus (3.84 log CFU/cm2). Furthermore, a combined concentration of 850 ppm Ag-NPs and 400 ppm ZnO-NPs showed high antimicrobial efficacy against E. coli (5.80 log CFU/cm2) and S. aureus (4.11 log CFU/cm2). The results also showed a high correlation between concentration levels and the bacterial activity of Ag–ZnO-NPs (R2 = 0.97 for S. aureus, and R2 = 0.99 for E. coli). They also showed that unlike individual action, the joint action of Ag-NPs and ZnO-NPs has high antimicrobial efficacy for both types of microorganisms. Moreover, Ag-NPs prevent the biofilm formation of L. monocytogenes in humid conditions of growth at concentrations of 500 ppm. Additional studies under different conditions are needed to test the durability of nanoparticle containing polyester surfaces with antimicrobial properties to optimize their use.

scholarly journals Effect of Spermidine on Biofilm Formation in Escherichia coli K-12

2021 ◽  
Vol 203 (10) ◽  
Author(s):  
Kullathida Thongbhubate ◽  
Yuko Nakafuji ◽  
Rina Matsuoka ◽  
Sonomi Kakegawa ◽  
Hideyuki Suzuki
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Conclusive Evidence ◽  
Bacterial Biofilm ◽  
Spermidine Synthase ◽  
Periplasmic Binding Protein ◽  
Content Type ◽  
E Coli ◽  
K 12

ABSTRACT Polyamines are essential for biofilm formation in Escherichia coli, but it is still unclear which polyamines are primarily responsible for this phenomenon. To address this issue, we constructed a series of E. coli K-12 strains with mutations in genes required for the synthesis and metabolism of polyamines. Disruption of the spermidine synthase gene (speE) caused a severe defect in biofilm formation. This defect was rescued by the addition of spermidine to the medium but not by putrescine or cadaverine. A multidrug/spermidine efflux pump membrane subunit (MdtJ)-deficient strain was anticipated to accumulate more spermidine and result in enhanced biofilm formation compared to the MdtJ+ strain. However, the mdtJ mutation did not affect intracellular spermidine or biofilm concentrations. E. coli has the spermidine acetyltransferase (SpeG) and glutathionylspermidine synthetase/amidase (Gss) to metabolize intracellular spermidine. Under biofilm-forming conditions, not Gss but SpeG plays a major role in decreasing the too-high intracellular spermidine concentrations. Additionally, PotFGHI can function as a compensatory importer of spermidine when PotABCD is absent under biofilm-forming conditions. Last, we report here that, in addition to intracellular spermidine, the periplasmic binding protein (PotD) of the spermidine preferential ABC transporter is essential for stimulating biofilm formation. IMPORTANCE Previous reports have speculated on the effect of polyamines on bacterial biofilm formation. However, the regulation of biofilm formation by polyamines in Escherichia coli has not yet been assessed. The identification of polyamines that stimulate biofilm formation is important for developing novel therapies for biofilm-forming pathogens. This study sheds light on biofilm regulation in E. coli. Our findings provide conclusive evidence that only spermidine can stimulate biofilm formation in E. coli cells, not putrescine or cadaverine. Last, ΔpotD inhibits biofilm formation even though the spermidine is synthesized inside the cells from putrescine. Since PotD is significant for biofilm formation and there is no ortholog of the PotABCD transporter in humans, PotD could be a target for the development of biofilm inhibitors.

scholarly journals Genotypic and Phenotypic Characteristics Associated with Biofilm Formation by Human Clinical Escherichia coli Isolates of Different Pathotypes

2017 ◽  
Vol 83 (24) ◽  
Author(s):  
Juliane Schiebel ◽  
Alexander Böhm ◽  
Jörg Nitschke ◽  
Michał Burdukiewicz ◽  
Jörg Weinreich ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Polymeric Matrix ◽  
Bacterial Biofilm ◽  
Automated Image Analysis ◽  
Host Immune System ◽  
Content Type ◽  
E Coli ◽  
Initial Adhesion ◽  
K 12

ABSTRACT Bacterial biofilm formation is a widespread phenomenon and a complex process requiring a set of genes facilitating the initial adhesion, maturation, and production of the extracellular polymeric matrix and subsequent dispersal of bacteria. Most studies on Escherichia coli biofilm formation have investigated nonpathogenic E. coli K-12 strains. Due to the extensive focus on laboratory strains in most studies, there is poor information regarding biofilm formation by pathogenic E. coli isolates. In this study, we genotypically and phenotypically characterized 187 human clinical E. coli isolates representing various pathotypes (e.g., uropathogenic, enteropathogenic, and enteroaggregative E. coli). We investigated the presence of biofilm-associated genes (“genotype”) and phenotypically analyzed the isolates for motility and curli and cellulose production (“phenotype”). We developed a new screening method to examine the in vitro biofilm formation ability. In summary, we found a high prevalence of biofilm-associated genes. However, we could not detect a biofilm-associated gene or specific phenotype correlating with the biofilm formation ability. In contrast, we did identify an association of increased biofilm formation with a specific E. coli pathotype. Enteroaggregative E. coli (EAEC) was found to exhibit the highest capacity for biofilm formation. Using our image-based technology for the screening of biofilm formation, we demonstrated the characteristic biofilm formation pattern of EAEC, consisting of thick bacterial aggregates. In summary, our results highlight the fact that biofilm-promoting factors shown to be critical for biofilm formation in nonpathogenic strains do not reflect their impact in clinical isolates and that the ability of biofilm formation is a defined characteristic of EAEC. IMPORTANCE Bacterial biofilms are ubiquitous and consist of sessile bacterial cells surrounded by a self-produced extracellular polymeric matrix. They cause chronic and device-related infections due to their high resistance to antibiotics and the host immune system. In nonpathogenic Escherichia coli, cell surface components playing a pivotal role in biofilm formation are well known. In contrast, there is poor information for their role in biofilm formation of pathogenic isolates. Our study provides insights into the correlation of biofilm-associated genes or specific phenotypes with the biofilm formation ability of commensal and pathogenic E. coli. Additionally, we describe a newly developed method enabling qualitative biofilm analysis by automated image analysis, which is beneficial for high-throughput screenings. Our results help to establish a better understanding of E. coli biofilm formation.

scholarly journals Antimicrobial effects of airborne acoustic ultrasound and plasma activated water from cold and thermal plasma systems on biofilms

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Clémentine M. G. Charoux ◽  
Apurva D. Patange ◽  
Laura M. Hinds ◽  
Jeremy C. Simpson ◽  
Colm P. O’Donnell ◽  
...  
Keyword(s):  
Thermal Plasma ◽  
Biofilm Formation ◽  
Synergistic Effects ◽  
Antimicrobial Properties ◽  
Bacterial Count ◽  
Bacterial Biofilm ◽  
Antimicrobial Effects ◽  
E Coli ◽  
And Control ◽  
Plasma Activated Water

Abstract Bacterial biofilms are difficult to inactivate due to their high antimicrobial resistance. Therefore, new approaches are required for more effective bacterial biofilm inactivation. Airborne acoustic ultrasound improves bactericidal or bacteriostatic activity which is safe and environmentally friendly. While, plasma activated water (PAW) is attracting increasing attention due to its strong antimicrobial properties. This study determined efficacy of combined airborne acoustic ultrasound and plasma activated water from both cold and thermal plasma systems in inactivating Escherichia coli K12 biofilms. The application of airborne acoustic ultrasound (15 min) alone was significantly more effective in reducing E. coli counts in 48 and 72 h biofilms compared to 30 min treatment with PAW. The effect of airborne acoustic ultrasound was more pronounced when used in combination with PAW. Airborne acoustic ultrasound treatment for 15 min of the E. coli biofilm followed by treatment with PAW significantly reduced the bacterial count by 2.2—2.62 Log10 CFU/mL when compared to control biofilm treated with distilled water. This study demonstrates that the synergistic effects of airborne acoustic ultrasound and PAW for enhanced antimicrobial effects. These technologies have the potential to prevent and control biofilm formation in food and bio-medical applications.

scholarly journals Magneto-mechanically actuated microstructures to efficiently prevent bacterial biofilm formation

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
S. Leulmi Pichot ◽  
H. Joisten ◽  
A. J. Grant ◽  
B. Dieny ◽  
R. P. Cowburn
Keyword(s):  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Substrate Surface ◽  
Low Frequency ◽  
Initial Position ◽  
Bacterial Biofilm ◽  
Biofilm Growth ◽  
E Coli ◽  
Initial Stage ◽  
Fouling Release

Abstract Biofilm colonisation of surfaces is of critical importance in various areas ranging from indwelling medical devices to industrial setups. Of particular importance is the reduced susceptibility of bacteria embedded in a biofilm to existing antimicrobial agents. In this paper, we demonstrate that remotely actuated magnetic cantilevers grafted on a substrate act efficiently in preventing bacterial biofilm formation. When exposed to an alternating magnetic field, the flexible magnetic cantilevers vertically deflect from their initial position periodically, with an extremely low frequency (0.16 Hz). The cantilevers’ beating prevents the initial stage of bacterial adhesion to the substrate surface and the subsequent biofilm growth. Our experimental data on E. coli liquid cultures demonstrate up to a 70% reduction in biofilm formation. A theoretical model has been developed to predict the amplitude of the cantilevers vertical deflection. Our results demonstrate proof-of-concept for a device that can magneto-mechanically prevent the first stage in bacterial biofilm formation, acting as on-demand fouling release active surfaces.

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