scholarly journals Identification of Biofilm Matrix-Associated Proteins from an Acid Mine Drainage Microbial Community

2011 ◽  
Vol 77 (15) ◽  
pp. 5230-5237 ◽  
Author(s):  
Yongqin Jiao ◽  
Patrik D'haeseleer ◽  
Brian D. Dill ◽  
Manesh Shah ◽  
Nathan C. VerBerkmoes ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Spatial Organization ◽  
Extracellular Polymeric Substances ◽  
Developmental Stages ◽  
Mine Drainage ◽  
Cell Envelope ◽  
Shotgun Proteomics ◽  
Extracellular Proteins ◽  
Cold Shock Protein ◽  
Biofilm Development

ABSTRACTIn microbial communities, extracellular polymeric substances (EPS), also called the extracellular matrix, provide the spatial organization and structural stability during biofilm development. One of the major components of EPS is protein, but it is not clear what specific functions these proteins contribute to the extracellular matrix or to microbial physiology. To investigate this in biofilms from an extremely acidic environment, we used shotgun proteomics analyses to identify proteins associated with EPS in biofilms at two developmental stages, designated DS1 and DS2. The proteome composition of the EPS was significantly different from that of the cell fraction, with more than 80% of the cellular proteins underrepresented or undetectable in EPS. In contrast, predicted periplasmic, outer membrane, and extracellular proteins were overrepresented by 3- to 7-fold in EPS. Also, EPS proteins were more basic by ∼2 pH units on average and about half the length. When categorized by predicted function, proteins involved in motility, defense, cell envelope, and unknown functions were enriched in EPS. Chaperones, such as histone-like DNA binding protein and cold shock protein, were overrepresented in EPS. Enzymes, such as protein peptidases, disulfide-isomerases, and those associated with cell wall and polysaccharide metabolism, were also detected. Two of these enzymes, identified as β-N-acetylhexosaminidase and cellulase, were confirmed in the EPS fraction by enzymatic activity assays. Compared to the differences between EPS and cellular fractions, the relative differences in the EPS proteomes between DS1 and DS2 were smaller and consistent with expected physiological changes during biofilm development.

scholarly journals Respiratory Heterogeneity Shapes Biofilm Formation and Host Colonization in Uropathogenic Escherichia coli

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Connor J. Beebout ◽  
Allison R. Eberly ◽  
Sabrina H. Werby ◽  
Seth A. Reasoner ◽  
John R. Brannon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Bacterial Communities ◽  
Spatial Organization ◽  
Bet Hedging ◽  
Content Type ◽  
Dna Organization ◽  
Quinol Oxidases ◽  
Biofilm Development ◽  
Biofilm Community

ABSTRACT Biofilms are multicellular bacterial communities encased in a self-secreted extracellular matrix comprised of polysaccharides, proteinaceous fibers, and DNA. Organization of these components lends spatial organization to the biofilm community such that biofilm residents can benefit from the production of common goods while being protected from exogenous insults. Spatial organization is driven by the presence of chemical gradients, such as oxygen. Here we show that two quinol oxidases found in Escherichia coli and other bacteria organize along the biofilm oxygen gradient and that this spatially coordinated expression controls architectural integrity. Cytochrome bd, a high-affinity quinol oxidase required for aerobic respiration under hypoxic conditions, is the most abundantly expressed respiratory complex in the biofilm community. Depletion of the cytochrome bd-expressing subpopulation compromises biofilm complexity by reducing the abundance of secreted extracellular matrix as well as increasing cellular sensitivity to exogenous stresses. Interrogation of the distribution of quinol oxidases in the planktonic state revealed that ∼15% of the population expresses cytochrome bd at atmospheric oxygen concentration, and this population dominates during acute urinary tract infection. These data point toward a bet-hedging mechanism in which heterogeneous expression of respiratory complexes ensures respiratory plasticity of E. coli across diverse host niches. IMPORTANCE Biofilms are multicellular bacterial communities encased in a self-secreted extracellular matrix comprised of polysaccharides, proteinaceous fibers, and DNA. Organization of these components lends spatial organization in the biofilm community. Here we demonstrate that oxygen gradients in uropathogenic Escherichia coli (UPEC) biofilms lead to spatially distinct expression programs for quinol oxidases—components of the terminal electron transport chain. Our studies reveal that the cytochrome bd-expressing subpopulation is critical for biofilm development and matrix production. In addition, we show that quinol oxidases are heterogeneously expressed in planktonic populations and that this respiratory heterogeneity provides a fitness advantage during infection. These studies define the contributions of quinol oxidases to biofilm physiology and suggest the presence of respiratory bet-hedging behavior in UPEC.

The Abundance and Organization of Salmonella Extracellular Polymeric Substances in Gallbladder-Mimicking Environments and In Vivo

2021 ◽  
Author(s):  
Mark M. Hahn ◽  
Juan F. González ◽  
Regan Hitt ◽  
Lauren Tucker ◽  
John S. Gunn
Keyword(s):  
Spatial Organization ◽  
Extracellular Polymeric Substances ◽  
Wild Type ◽  
Chronic Infections ◽  
Human Bile ◽  
Cholesterol Gallstones ◽  
Coated Surfaces ◽  
Biofilm Development

Salmonella enterica serovar Typhi ( S. Typhi ) causes chronic infections by establishing biofilms on cholesterol gallstones. Production of extracellular polymeric substances (EPSs) is key to biofilm development and biofilm architecture depends on which EPSs are made. The presence and spatial distribution of Salmonella EPSs produced in vitro and in vivo were investigated in S. Typhi murium and S. Typhi biofilms by confocal microscopy. Comparisons between serovars and EPS-mutant bacteria were examined by growth on cholesterol-coated surfaces, with human gallstones in ox or human bile, and in mice with gallstones. On cholesterol-coated surfaces, major differences in EPS biomass were not found between serovars. Co-culture biofilms containing wild-type (WT) and EPS-mutant bacteria demonstrated WT compensation for EPS mutations. Biofilm EPS analysis from gallbladder-mimicking conditions found that culture in human bile more consistently replicated the relative abundance and spatial organization of each EPS on gallstones from the chronic mouse model than culture in ox bile. S. Typhi murium biofilms cultured in vitro on gallstones in ox bile exhibited co-localized pairings of curli fimbriae/lipopolysaccharide and O antigen capsule/cellulose while these associations were not present in S. Typhi biofilms or in mouse gallstone biofilms. In general, inclusion of human bile with gallstones in vitro replicated biofilm development on gallstones in vivo , demonstrating its strength as a model for studying biofilm parameters or EPS-directed therapeutic treatments.

scholarly journals Streptococcus mutans yidC1andyidC2Impact Cell Envelope Biogenesis, the Biofilm Matrix, and Biofilm Biophysical Properties

2018 ◽  
Vol 201 (1) ◽  
Author(s):  
Sara R. Palmer ◽  
Zhi Ren ◽  
Geelsu Hwang ◽  
Yuan Liu ◽  
Ashton Combs ◽  
...  
Keyword(s):  
Physical Properties ◽  
Streptococcus Mutans ◽  
Spatial Organization ◽  
Mechanical Stability ◽  
Cell Envelope ◽  
Wild Type ◽  
Biophysical Properties ◽  
Content Type ◽  
Glucan Synthesis ◽  
Biofilm Development

ABSTRACTProper envelope biogenesis ofStreptococcus mutans, a biofilm-forming and dental caries-causing oral pathogen, requires two paralogs (yidC1andyidC2) of the universally conserved YidC/Oxa1/Alb3 family of membrane integral chaperones and insertases. The deletion of either paralog attenuates virulencein vivo, but the mechanisms of disruption remain unclear. Here, we determined whether the deletion ofyidCaffects cell surface properties, extracellular glucan production, and/or the structural organization of the exopolysaccharide (EPS) matrix and biophysical properties ofS. mutansbiofilm. Compared to the wild type, the ΔyidC2 mutant lacked staining with fluorescent vancomycin at the division septum, while the ΔyidC1mutant resembled the wild type. Additionally, the deletion of eitheryidC1oryidC2resulted in less insoluble glucan synthesis but produced more soluble glucans, especially at early and mid-exponential-growth phases. Alteration of glucan synthesis by both mutants yielded biofilms with less dry weight and insoluble EPS. In particular, the deletion ofyidC2resulted in a significant reduction in biofilm biomass and pronounced defects in the spatial organization of the EPS matrix, thus modifying the three-dimensional (3D) biofilm architecture. The defective biofilm harbored smaller bacterial clusters with high cell density and less surrounding EPS than those of the wild type, which was stiffer in compression yet more susceptible to removal by shear. Together, our results indicate that the elimination of eitheryidCparalog results in changes to the cell envelope and glucan production that ultimately disrupts biofilm development and EPS matrix structure/composition, thereby altering the physical properties of the biofilms and facilitating their removal. YidC proteins, therefore, represent potential therapeutic targets for cariogenic biofilm control.IMPORTANCEYidC proteins are membrane-localized chaperone insertases that are universally conserved in all bacteria and are traditionally studied in the context of membrane protein insertion and assembly. Both YidC paralogs of the cariogenic pathogenStreptococcus mutansare required for proper envelope biogenesis and full virulence, indicating that these proteins may also contribute to optimal biofilm formation in streptococci. Here, we show that the deletion of eitheryidCresults in changes to the structure and physical properties of the EPS matrix produced byS. mutans, ultimately impairing optimal biofilm development, diminishing its mechanical stability, and facilitating its removal. Importantly, the universal conservation of bacterialyidCorthologs, combined with our findings, provide a rationale for YidC as a possible drug target for antibiofilm therapies.

scholarly journals Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure

mBio ◽  
2013 ◽  
Vol 4 (5) ◽  
Author(s):  
Chia Hung ◽  
Yizhou Zhou ◽  
Jerome S. Pinkner ◽  
Karen W. Dodson ◽  
Jan R. Crowley ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Biofilm Formation ◽  
Developmental Stages ◽  
Three Dimensional ◽  
Bacterial Biofilms ◽  
Content Type ◽  
Abiotic Surfaces ◽  
Biochemical Analyses ◽  
Biofilm Development

ABSTRACTBacterial biofilms are ubiquitous in nature, and their resilience is derived in part from a complex extracellular matrix that can be tailored to meet environmental demands. Although common developmental stages leading to biofilm formation have been described, how the extracellular components are organized to allow three-dimensional biofilm development is not well understood. Here we show that uropathogenicEscherichia coli(UPEC) strains produce a biofilm with a highly ordered and complex extracellular matrix (ECM). We used electron microscopy (EM) techniques to image floating biofilms (pellicles) formed by UPEC. EM revealed intricately constructed substructures within the ECM that encase individual, spatially segregated bacteria with a distinctive morphology. Mutational and biochemical analyses of these biofilms confirmed curli as a major matrix component and revealed important roles for cellulose, flagella, and type 1 pili in pellicle integrity and ECM infrastructure. Collectively, the findings of this study elucidated that UPEC pellicles have a highly organized ultrastructure that varies spatially across the multicellular community.IMPORTANCEBacteria can form biofilms in diverse niches, including abiotic surfaces, living cells, and at the air-liquid interface of liquid media. Encasing these cellular communities is a self-produced extracellular matrix (ECM) that can be composed of proteins, polysaccharides, and nucleic acids. The ECM protects biofilm bacteria from environmental insults and also makes the dissolution of biofilms very challenging. As a result, formation of biofilms within humans (during infection) or on industrial material (such as water pipes) has detrimental and costly effects. In order to combat bacterial biofilms, a better understanding of components required for biofilm formation and the ECM is required. This study defined the ECM composition and architecture of floating pellicle biofilms formed byEscherichia coli.

scholarly journals The Role of Exopolysaccharides in Oral Biofilms

2019 ◽  
Vol 98 (7) ◽  
pp. 739-745 ◽  
Author(s):  
C. Cugini ◽  
M. Shanmugam ◽  
N. Landge ◽  
N. Ramasubbu
Keyword(s):  
Biofilm Formation ◽  
Bacterial Colonization ◽  
Super Resolution ◽  
Extracellular Proteins ◽  
Extracellular Polysaccharides ◽  
Oral Biofilms ◽  
The Matrix ◽  
Oral Microbes ◽  
Biofilm Development

The oral cavity contains a rich consortium of exopolysaccharide-producing microbes. These extracellular polysaccharides comprise a major component of the oral biofilm. Together with extracellular proteins, DNA, and lipids, they form the biofilm matrix, which contributes to bacterial colonization, biofilm formation and maintenance, and pathogenesis. While a number of oral microbes have been studied in detail with regard to biofilm formation and pathogenesis, the exopolysaccharides have been well characterized for only select organisms, namely Streptococcus mutans and Aggregatibacter actinomycetemcomitans. Studies on the exopolysaccharides of other oral organisms, however, are in their infancy. In this review, we present the current research on exopolysaccharides of oral microbes regarding their biosynthesis, regulation, contributions to biofilm formation and stability of the matrix, and immune evasion. In addition, insight into the role of exopolysaccharides in biofilms is highlighted through the evaluation of emerging techniques such as pH probing of biofilm colonies, solid-state nuclear magnetic resonance for macromolecular interactions within biofilms, and super-resolution microscopy analysis of biofilm development. Finally, exopolysaccharide as a potential nutrient source for species within a biofilm is discussed.

Abstract MP254: Decellularized Extracellular Matrix Hydrogel Lowers Fibrosis And Fibroblast Differentiation In Post-injury Mice Hearts Through Capg

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Xinming Wang ◽  
Samuel Senyo
Keyword(s):  
Extracellular Matrix ◽  
Molecular Mechanisms ◽  
Smooth Muscle Actin ◽  
Growth Factor Receptor ◽  
Inhibitory Effect ◽  
Extracellular Proteins ◽  
Collagen Deposition ◽  
Post Injury ◽  
Capping Protein ◽  
Decellularized Extracellular Matrix

Hypothesis and objective: We hypothesize that transplantation of decellularized cardiac extracellular matrix (dECM) lowers fibrosis and fibroblast differentiation. In this study we investigated collagen deposition and fibroblast differentiation in post-MI hearts and heart explants of various stiffness after dECM hydrogel treatments. The objectives are 1) determining if dECM derived from fetal and adult porcine hearts reduces fibrosis in injured hearts; and 2) identifying specific signaling pathways that regulate fibroblasts differentiation induced by extracellular proteins. Methods: Porcine dECM was injected immediately after ligating coronary artery in P1 mice. Histology was conducted on day 7 post-myocardial infarction (MI). A mice ventricle explant model was used to investigate the molecular mechanisms. Results: We observed that fetal dECM treatment lowered fibrosis and fibroblast differentiation in post-MI hearts (Fig.1). Fibroblast differentiation as indicated by α-smooth muscle actin expression in vimentin or platelet derived growth factor receptor α positive cells showed an inhibitory effect of fetal dECM on fibroblast differentiation. Using a heart explant model of modulated microenvironment stiffness, we demonstrated that increasing tissue stiffness stimulates fibroblast differentiation and collagen deposition. Fetal dECM treatment, however, inhibited fibroblast differentiation induced by increasing microenvironment stiffness. Transcriptome analysis revealed that two cytoskeleton-related genes, macrophage capping protein (CAPG) and leupaxin (LPXN), are modulated by dECM treatments. Using cytoskeleton polymerization modulators and siRNA, we demonstrated that fetal dECM lowers fibroblast differentiation through CAPG.

scholarly journals Lon Protease Has Multifaceted Biological Functions inAcinetobacter baumannii

2018 ◽  
Vol 201 (2) ◽  
Author(s):  
Carly Ching ◽  
Brendan Yang ◽  
Chineme Onwubueke ◽  
David Lazinski ◽  
Andrew Camilli ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Developmental Process ◽  
Cell Envelope ◽  
Opportunistic Pathogen ◽  
Lon Protease ◽  
Content Type ◽  
Hospital Acquired Infections ◽  
Biofilm Development ◽  
Hospital Acquired

ABSTRACTAcinetobacter baumanniiis a Gram-negative opportunistic pathogen that is known to survive harsh environmental conditions and is a leading cause of hospital-acquired infections. Specifically, multicellular communities (known as biofilms) ofA. baumanniican withstand desiccation and survive on hospital surfaces and equipment. Biofilms are bacteria embedded in a self-produced extracellular matrix composed of proteins, sugars, and/or DNA. Bacteria in a biofilm are protected from environmental stresses, including antibiotics, which provides the bacteria with selective advantage for survival. Although some gene products are known to play roles in this developmental process inA. baumannii, mechanisms and signaling remain mostly unknown. Here, we find that Lon protease inA. baumanniiaffects biofilm development and has other important physiological roles, including motility and the cell envelope. Lon proteases are found in all domains of life, participating in regulatory processes and maintaining cellular homeostasis. These data reveal the importance of Lon protease in influencing keyA. baumanniiprocesses to survive stress and to maintain viability.IMPORTANCEAcinetobacter baumanniiis an opportunistic pathogen and is a leading cause of hospital-acquired infections.A. baumanniiis difficult to eradicate and to manage, because this bacterium is known to robustly survive desiccation and to quickly gain antibiotic resistance. We sought to investigate biofilm formation inA. baumannii, since much remains unknown about biofilm formation in this bacterium. Biofilms, which are multicellular communities of bacteria, are surface attached and difficult to eliminate from hospital equipment and implanted devices. Our research identifies multifaceted physiological roles for the conserved bacterial protease Lon inA. baumannii. These roles include biofilm formation, motility, and viability. This work broadly affects and expands understanding of the biology ofA. baumannii, which will permit us to find effective ways to eliminate the bacterium.

scholarly journals Bacterial Cellulose from Food to Biomedical Products

2020 ◽  
Vol 14 (1) ◽  
pp. 124-133
Author(s):  
Supajit Sraphet ◽  
Bagher Javadi
Keyword(s):  
Bacterial Cellulose ◽  
Extracellular Polymeric Substances ◽  
Control Strategies ◽  
Low Cost ◽  
Cell Communication ◽  
Future Application ◽  
Membrane Structures ◽  
Food Ingredient ◽  
Expression Of Genes ◽  
Biofilm Development

Cellulose production of aerobic bacteria with its very unique physiochemical properties attracted many researchers. The biosynthetic of Bacterial Cellulose (BC) was produced by low-cost media recently. BC has been used as biomaterials and food ingredient these days. Moreover, the capacity of BC composite gives the numerous application opportunities in other fields. Bacterial Cellulose (BC) development is differentiated from suspension planktonic culture by their Extracellular Polymeric Substances (EPS), down-regulation of growth rate and up-down the expression of genes. The attachment of microorganisms is highly dependent on their cell membrane structures and growth medium. This is a very complicated phenomenon that optimal conditions defined the specific architecture. This architecture is made of microbial cells and EPS. Cell growth and cell communication mechanisms effect biofilm development and detachment. Understandings of development and architecture mechanisms and control strategies have a great impact on the management of BC formation with beneficial microorganisms. This mini-review paper presents the overview of outstanding findings from isolating and characterizing the diversity of bacteria to BC's future application, from food to biosensor products. The review would help future researchers in the sustainable production of BC, applications advantages and opportunities in food industry, biomaterial and biomedicine.

Effects of extracellular polymeric substances on granulation of anoxic sludge in sequencing batch reactor

2012 ◽  
Vol 66 (3) ◽  
pp. 543-548
Author(s):  
Binbin Wang ◽  
Shunlian Liu ◽  
Hongmei Zhao ◽  
Xinyan Zhang ◽  
Dangcong Peng
Keyword(s):  
Extracellular Polymeric Substances ◽  
Sequencing Batch Reactor ◽  
Batch Reactor ◽  
Cation Exchange Resin ◽  
Granular Sludge ◽  
Extracellular Proteins ◽  
Extracellular Polysaccharides ◽  
Positive Role ◽  
Start Up ◽  
Sludge Granulation

Variations of extracellular polymeric substances (EPS) and its components with sludge granulation were examined in a lab-scale sequencing batch reactor (SBR) which was fed with sodium nitrate and sodium acetate. Ultrasonication plus cation exchange resin (CER) were used as the EPS extraction method. Results showed that after approximately 90 d cultivation, the sludge in the reactor was almost granulated. The content of extracellular polysaccharides increased from 10.36 mg/g-VSS (volatile suspended solids) at start-up with flocculent sludge to 23.18 mg/g-VSS at 91 d with matured granular sludge, while the content of extracellular proteins were almost unchanged. Polysaccharides were the major components of EPS in anoxic granular sludge, accounting for about 70.6–79.0%, while proteins and DNA accounted for about 16.5–18.9% and 4.6–9.9%, respectively. It is proposed that EPS play a positive role in anoxic sludge granulation and polysaccharides might be strongly involved in aggregation of flocs into granules.

scholarly journals Characterization of Temporal Protein Production in Pseudomonas aeruginosa Biofilms

2005 ◽  
Vol 187 (23) ◽  
pp. 8114-8126 ◽  
Author(s):  
Christopher J. Southey-Pillig ◽  
David G. Davies ◽  
Karin Sauer
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Biofilm Formation ◽  
Protein Production ◽  
Developmental Stages ◽  
Developmental Process ◽  
Developmental Cycle ◽  
Specific Proteins ◽  
Biofilm Development ◽  
The Impact

ABSTRACT Phenotypic and genetic evidence supporting the notion of biofilm formation as a developmental process is growing. In the present work, we provide additional support for this hypothesis by identifying the onset of accumulation of biofilm-stage specific proteins during Pseudomonas aeruginosa biofilm maturation and by tracking the abundance of these proteins in planktonic and three biofilm developmental stages. The onset of protein production was found to correlate with the progression of biofilms in developmental stages. Protein identification revealed that proteins with similar function grouped within similar protein abundance patterns. Metabolic and housekeeping proteins were found to group within a pattern separate from virulence, antibiotic resistance, and quorum-sensing-related proteins. The latter were produced in a progressive manner, indicating that attendant features that are characteristic of biofilms such as antibiotic resistance and virulence may be part of the biofilm developmental process. Mutations in genes for selected proteins from several protein production patterns were made, and the impact of these mutations on biofilm development was evaluated. The proteins cytochrome c oxidase, a probable chemotaxis transducer, a two-component response regulator, and MexH were produced only in mature and late-stage biofilms. Mutations in the genes encoding these proteins did not confer defects in growth, initial attachment, early biofilm formation, or twitching motility but were observed to arrest biofilm development at the stage of cell cluster formation we call the maturation-1 stage. The results indicated that expression of theses genes was required for the progression of biofilms into three-dimensional structures on abiotic surfaces and the completion of the biofilm developmental cycle. Reverse transcription-PCR analysis confirmed the detectable change in expression of the respective genes ccoO, PA4101, and PA4208. We propose a possible mechanism for the role of these biofilm-specific proteins in biofilm formation.

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