Increase In Cell Surface Hydrophobicity Research Articles

Inhibition of the adhesion to catheters of uropathogenic Escherichia coli by sub-inhibitory concentrations of cefotaxime

ObjectiveTo investigate the effect of sub-inhibitory concentrations of cefotaxime on adherence to siliconised latex urinary catheters of uropathogenic Escherichia coli strains from pregnant and non pregnant patients. Study designUsing random sampling, 30 E. coli strains were selected from hospitalized patients with catheter associated urinary tract infection, 12 from pregnant women and 18 from men and non-pregnant women. The strains were categorized on the basis of cefotaxime susceptibility, adhesion and biofilm production capacity, cell surface hydrophobicity and expression of adhesins and fimbriae in vitro. ResultsThe overall results indicated that sub-inhibitory concentrations of cefotaxime could reduce the adhesiveness, the biofilm production and hence, potentially, the infection rate associated with indwelling urinary catheters. ConclusionBased on our results, we propose that this reduction is due to decreasing exopolysaccharide production and increasing cell surface hydrophobicity of E.coli strains.

Long-term influence of the presence of a non-aqueous phase on the cell surface hydrophobicity of Pseudomonas in two-phase partitioning bioreactors

The long-term influence of silicone oil 200 cSt (SO200) and 2, 2, 4, 4, 6, 8, 8-heptamethylnonane (HMN) on the cell surface hydrophobicity (CSH) of a hexane-degrading Pseudomonas aeruginosa strain and a toluene-degrading Pseudomonas putida strain was assessed in two-phase partitioning bioreactors under batch and continuous operation. CSH was evaluated using a modified BATH method based on optical density (CSH(OD)) and colony-forming unit (CSH(CFU)) measurements. In the presence of HMN, P. aeruginosa turned hydrophobic over the time course as shown by the gradual increase in CSH(OD) (61 ± 1%) and CSH(CFU) (53 ± 3%) under batch degradation and in CSH(OD) (49 ± 0%) under continuous operation. However, P. putida turned hydrophobic only under continuous operation ([Formula: see text]). On the other hand, no significant CSH enhancement was observed in both Pseudomonas strains in the presence of SO200. These results suggested that CSH is species, non-aqueous phase, and cultivation mode dependant, and an inducible property of bacteria. Maximum hexane elimination capacities increased by 2 and 3 in the presence of SO200 and HMN, respectively. Based on the absence of CSH in P. aeruginosa in the presence of SO200, the higher elimination capacities recorded were likely due to an improved hexane mass transfer (physical effect). However, in the presence of HMN, a direct hexane uptake from the non-aqueous phase (biological effect) might have also contributed to this enhancement.

FLO11 is essential for pellicle formation by wild pellicle-forming yeasts isolated from contaminated wines

The Δflo11/Δflo11 disruptant strains constructed from three wild yeast strains, which were independently isolated from pellicles formed on the surface of contaminated stored wines, completely lost their ability to form pellicles. The FLO11 expression level of the pellicle-forming strains was much higher than that of non-pellicle-forming wine strains. Cell surface hydrophobicity of the Δflo11/Δflo11 disruptant strains was as high as that of the parental pellicle-forming strains. These findings indicate that FLO11 is essential for unfavorable pellicle formation of wild pellicle-forming yeasts by mechanisms other than increasing cell surface hydrophobicity.

Surface hydrophobicity of petroleum hydrocarbon degrading Burkholderia strains and their interactions with NAPLs and surfaces

Bacterial cell surface hydrophobicity (CSH) is an important factor governing the growth and adhesion behavior of microorganisms on non-aqueous phase liquids (NAPLs). In this work CSH and surface charge was quantified for three oil degrading Burkholderia cultures: aliphatic degrader Burkholderia cepacia (ES1) and two strains of aromatic degrading Burkholderia multivorans (NG1 and HN1) based on contact angle and zeta potential measurement. Model non-aqueous phase liquids (NAPLs) were formulated using n-hexadecane, naphthalene, phenanthrene and pyrene in varying concentration. Adhesion on to glass surfaces of varying hydrophobicity and adherence to n-hexadecane was quantified and correlated with hydrophobicity of the surface; variation in CSH of the culture in response to model NAPL used as growth substrate; and variation in zeta potential as a result of variation in growth substrate, ionic strength and pH of resuspension solution. B. cepacia (ES1) and B. multivorans (HN1) depicted comparable CSH which was higher than that of B. multivorans (NG1). For each culture, CSH was found to vary with the model NAPL used as growth substrate. Adhesion to glass increased with increase in CSH of the bacterial culture and with increase in hydrophobicity of the glass surface. B. cepacia (ES1) with lower negative zeta potential consistently depicted greater adhesion compared to B. multivorans (HN1). Adherence to n-hexadecane was significantly affected by various other factors, such as, growth substrate, pH, resuspension solution and their interactions as revealed through statistical analysis. These factors affected both the zeta potential and adherence to n-hexadecane to varying degree for the three Burkholderia cultures.

The Regulator RamA Influences cmytA Transcription and Cell Morphology of Corynebacterium ammoniagenes

RamA plays a regulatory role for acetate utilization and S-layer biosynthesis in Corynebacterium glutamicum. Looking for any additional role, the function of RamA was analyzed in Corynebacterium ammoniagenes, which is closely related to C. glutamicum. In this study, we showed that the DeltaramA mutant constructed by a markerless knockout strategy possessed increased cell surface hydrophobicity, leading to the formation of aggregated cell masses in liquid media. In addition, the mutant exhibited an elongated cell shape as observed by SEM, suggesting that cell wall-associated proteins might be influenced. Furthermore, cell surface proteome analysis revealed that the expression of cmytA gene encoding corynomycoloyl transferase required for cell wall biosynthesis was down-regulated in the mutant, supporting the regulatory role of RamA in cell wall assembly. These studies support a novel regulatory role of RamA in inducing the expression of proteins required for cell wall assembly.

Streptococcus mutans SMU.623c Codes for a Functional, Metal-Dependent Polysaccharide Deacetylase That Modulates Interactions with Salivary Agglutinin

The genome sequence of the oral pathogen Streptococcus mutans predicts the presence of two putative polysaccharide deacetylases. The first, designated PgdA in this paper, shows homology to the catalytic domains of peptidoglycan deacetylases from Streptococcus pneumoniae and Listeria monocytogenes, which are both thought to be involved in the bacterial defense mechanism against human mucosal lysozyme and are part of the CAZY family 4 carbohydrate esterases. S. mutans cells in which the pgdA gene was deleted displayed a different colony texture and a slightly increased cell surface hydrophobicity and yet did not become hypersensitive to lysozyme as shown previously for S. pneumoniae. To understand this apparent lack of activity, the high-resolution X-ray structure of S. mutans PgdA was determined; it showed the typical carbohydrate esterase 4 fold, with metal bound in a His-His-Asp triad. Analysis of the protein surface showed that an extended groove lined with aromatic residues is orientated toward the active-site residues. The protein exhibited metal-dependent de-N-acetylase activity toward a hexamer of N-acetylglucosamine. No activity was observed toward shorter chitooligosaccharides or a synthetic peptidoglycan tetrasaccharide. In agreement with the lysozyme data this would suggest that S. mutans PgdA does not act on peptidoglycan but on an as-yet-unidentified polysaccharide within the bacterial cell surface. Strikingly, the pgdA-knockout strain showed a significant increase in aggregation/agglutination by salivary agglutinin, in agreement with this gene acting as a deacetylase of a cell surface glycan.

Adsorption of monorhamnolipid and dirhamnolipid on two Pseudomonas aeruginosa strains and the effect on cell surface hydrophobicity

Previously, adsorption feature of a dirhamnolipid biosurfactant on diverse microbial cells was studied and the effect of the adsorption on cell surface hydrophobicity was compared. In this paper, the adsorption behavior of a monorhamnolipid and a dirhamnolipid on cells of two Pseudomonas aeruginosa strains was investigated in order to further reveal the influence of biosurfactant structure and cell property on the adsorption and the relation between the adsorption and cell surface hydrophobicity. Experimental results showed that the adsorption capacity of all the cells to monorhamnolipid was much stronger than to dirhamnolipid, and the rhamnolipid-sourced P. aeruginosa cells, no matter grown on glucose or hexadecane, released extra dirhamnolipid when aqueous concentration of dirhamnolipid was too high. Length of surfactant alkyl chain as well as the type of carbon source used to cultivate the cell adsorbents had only minor influence on the adsorption. The adsorption was assumed to be driven by polar interaction between the rhamnolipid molecules and the cell surface chemical groups. The directional orientation of the rhamnolipid molecules with hydrophobic moiety extending to the environment may account for the rapid increase of cell surface hydrophobicity at low aqueous concentrations of the surfactant, while the stable or decreased cell hydrophobicity was probably the consequence of multiple surfactant layer formation or hemimicelle accumulation.

AgfC and AgfE facilitate extracellular thin aggregative fimbriae synthesis in Salmonella Enteritidis

Salmonella thin aggregative fimbriae (Tafi; curli) are important in pathogenesis and biofilm formation; however, less is known of their structure and morphogenesis. In the Salmonella agfBAC Tafi operon, the transcription and role of agfC have been elusive. In this study, agfBAC transcripts were detected using a sensitive reverse transcriptase technique. Native AgfC was not detected using polyclonal antibodies generated against purified hexahistidine-tagged AgfC; however, in trans expression revealed that AgfC was localized to the periplasm as a mature form. An isogenic DeltaagfC mutant displayed an abundance of 20 nm fibres, in addition to native Tafi (5-7 nm), and had an increase in cell surface hydrophobicity. Purified 20 nm fibres were depolymerized under exceptionally stringent conditions to release what proved to be AgfA subunits. This revealed that the 20 nm fibres represented a different form of Tafi. The role of AgfC in Tafi assembly was investigated further using an antibody-capture assay of isogenic Deltaagf mutants. A soluble antibody-accessible form of AgfA was captured in wild-type (wt), DeltaagfB and DeltaagfF strains, in support of the extracellular nucleation-precipitation pathway of Tafi assembly, but not in DeltaagfC or DeltaagfE mutants. This indicates that AgfC and AgfE are important for AgfA extracellular assembly, facilitating the synthesis of Tafi.

Growth and physiological response of Arthrobacter protophormiae RKJ100 toward higher concentrations of o-nitrobenzoate and p-hydroxybenzoate

Bioremediation of sites that are heavily contaminated with pollutant chemicals is a challenge as most of the microorganisms cannot tolerate higher concentrations of toxic compounds. Only a few strains of the genus Pseudomonas have been studied for their tolerance toward the higher concentrations of aromatic pollutant compounds, a phenomenon that is accompanied by various physiological changes. In the present study we have characterized the growth response and physiological changes (adaptations) of a Gram-positive bacterium, Arthrobacter protophormiae RKJ100, toward the higher concentrations of two aromatic compounds, viz. o-nitrobenzoate (ONB) and p-hydroxybenzoate (PHB). Arthrobacter protophormiae RKJ100 could utilize 30 mM ONB and 50 mM PHB as sole sources of carbon and energy. It was capable of growth on higher concentrations of ONB (up to 200 mM) and PHB (up to 150 mM) when the cells were pre-exposed to lower concentrations of these compounds. The adaptive responses shown by the organism during growth on higher concentrations of these compounds were evident from significant changes in cellular fatty acid profiles. In addition, Bacterial Adhesion To Hydrocarbon (BATH) assay and scanning electron microscopy showed substantial increase in cell surface hydrophobicity and decrease in cell size of A. protophormiae RKJ100 when grown on ONB and PHB as compared to succinate-grown cells.

The role ofFLO11inSaccharomyces cerevisiaebiofilm development in a laboratory based flow-cell system

A role of the FLO11 in Saccharomyces cerevisiae biofilm development in a flow cell system was examined. We carried out an ectopic FLO11 expression in the wild type (wt) BY4741 strain that has low levels of endogenous FLO11 transcript. In contrast to the nonadhesive wt, the FLO11 overexpression strain (BY4741 FLO11(+)) readily adhered to both liquid-hydrophobic and liquid-hydrophilic solid interfaces and was able to grow as a biofilm monolayer in a flow system. Cellular features associated with FLO11 were examined and found to be consistent with the previous studies conducted in different strains of S. cerevisiae. When grown in suspended liquid culture, BY4741 FLO11(+) formed larger cellular aggregates (clumps), consisting of from five to 60 cells, and displayed an increased cell surface hydrophobicity, without changes in the cell size or growth rate, compared to wt. However, the invasive growth associated with FLO11 expression was not observed in BY4741 FLO11(+). The significance of these findings is discussed in the context of clinically and industrially relevant biofilms.

Characterization of bacterial isolates from industrial wastewater according to probable modes of hexadecane uptake

Bacterial isolates from industrial wastewater were characterized according to probable modes of hexadecane uptake based on data for cell surface hydrophobicity, emulsifying activity, glycoside content and surface tension of cell-free culture medium. The results obtained suggested that both modes of biosurfactant-enhanced hexadecane uptake by bacterial strains take place, direct uptake and alkane transfer. The increase in cell surface hydrophobicity and glycoside production by the strains suggested the existence of biosurfactant-enhanced interfacial uptake of the alkane. Such mechanism is probably predominant for three isolates, Staphylococcus sp. HW-2, Streptococcus sp. HW-9 and Bacillus sp. HW-4. Secreted biosurfactants enhanced mainly alkane emulsification for most hydrophobic isolate Arthrobacter sp. HW-8, and micellar transfer for most hydrophilic isolate Streptococcus sp. HW-5. For other strains (67%) both mechanisms of biosurfactant-enhanced hexadecane uptake probably take place in similar degree, interfacial uptake and alkane emulsification. The results obtained could contribute to clarifying the natural relationships between the members of water ecosystem studied as well as will reveal potential producers of surface active compounds.

Adaptive evolution by mutations in theFLO11gene

In nature, Saccharomyces yeasts manifest a number of adaptive responses to overcome adverse environments such as filamentation, invasive growth, flocculation and adherence to solid surfaces. Certain Saccharomyces wild yeasts, namely "flor yeasts," have also acquired the ability to form a buoyant biofilm at the broth surface. Here we report that mutations in a single gene, identified as FLO11, separate these "floating" yeasts from their nonfloating relatives. We have determined that the capability to form a self-supporting biofilm at the liquid surface is largely dependent on two changes in the FLO11 gene. First, we identified a 111-nt deletion within a repression region of the FLO11 promoter that significantly increases FLO11 gene expression. Secondly, we found rearrangements within the central tandem repeat domain of the coding region that yield a more hydrophobic Flo11p variant. Together, these mutations result in dramatic increase in cell surface hydrophobicity, which in turn confers these yeasts the ability to float by surface tension, an adaptive mechanism to gain direct access to oxygen within oxygen-poor liquid environments.

Effects of Punica granatum Pericarps and Quercus infectoria Nutgalls on Cell Surface Hydrophobicity and Cell Survival of Helicobacter pylori

The effect of ethanolic extracts of Punica granatum (P. granatum) and Quercus infectoria (Q. infectoria) on cell surface hydrophobicity of 10 clinically isolated Helicobacter pylori strains were investigated using salt aggregation test. Both P. granatum and Q. infectoria significantly increased the hydrophobicity of all isolates, irrespective of their antibiotic resistance patterns. These two plant species were demonstrated to produce both bacteriostatic and bactericidal activities. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were from 0.78 to 6.25 and 3.12 to 6.25 mg/ml for P. granatum, and from 3.12 to 6.25 and 3.12 to 12.5 mg/ml for Q. infectoria, respectively. Ethyl acetate and n-butanol fractions of both plants had values at least 10-fold lower than the MIC and MBC values of the ethanolic extracts. The results indicate no relationship of the increased cellsurface hydrophobicity with the MIC or MBC values.

The fungal hydrophobin RolA recruits polyesterase and laterally moves on hydrophobic surfaces

When fungi grow on plant or insect surfaces coated with wax polyesters that protect against pathogens, the fungi generally form aerial hyphae to contact the surfaces. Aerial structures such as hyphae and conidiophores are coated with hydrophobins, which are surface-active proteins involved in adhesion to hydrophobic surfaces. When the industrial fungus Aspergillus oryzae was cultivated in a liquid medium containing the biodegradable polyester polybutylene succinate-coadipate (PBSA), the rolA gene encoding hydrophobin RolA was highly transcribed. High levels of RolA and its localization on the cell surface in the presence of PBSA were confirmed by immunostaining. Under these conditions, A. oryzae simultaneously produced the cutinase CutL1, which hydrolyses PBSA. Pre-incubation of PBSA with RolA stimulated PBSA degradation by CutL1, suggesting that RolA bound to the PBSA surface was required for the stimulation. Immunostaining revealed that PBSA films coated with RolA specifically adsorbed CutL1. Quartz crystal microbalance analyses further demonstrated that RolA attached to a hydrophobic sensor chip specifically adsorbed CutL1. Circular dichroism spectra of soluble-state RolA and bound RolA suggested that RolA underwent a conformational change after its adsorption to hydrophobic surfaces. These results suggest that RolA adsorbed to the hydrophobic surface of PBSA recruits CutL1, resulting in condensation of CutL1 on the PBSA surface and consequent stimulation of PBSA hydrolysis. A fluorescence recovery after photobleaching experiment on PBSA films coated with FITC-labelled RolA suggested that RolA moves laterally on the film. We discuss the novel molecular functions of RolA with regard to plastic degradation.

FLO11-Based Model for Air-Liquid Interfacial Biofilm Formation bySaccharomyces cerevisiae

Sardinian wine strains of Saccharomyces cerevisiae used to make sherry-like wines form a biofilm at the air-liquid interface at the end of ethanolic fermentation, when grape sugar is depleted and further growth becomes dependent on access to oxygen. Here, we show that FLO11, which encodes a hydrophobic cell wall glycoprotein, is required for the air-liquid interfacial biofilm and that biofilm cells have a buoyant density greater than the suspending medium. We propose a model for biofilm formation based on an increase in cell surface hydrophobicity occurring at the diauxic shift. This increase leads to formation of multicellular aggregates that effectively entrap carbon dioxide, providing buoyancy. A visible biofilm appears when a sufficient number of hydrophobic cell aggregates are carried to and grow on the liquid surface.

Global cell surface conformational shift mediated by a Candida albicans adhesin.

Candida albicans maintains both commensal and pathogenic states in humans. Both states are dependent on cell surface-expressed adhesins, including those of the Als family. Heterologous expression of Als5p at the surface of Saccharomyces cerevisiae results in Als5p-mediated adhesion to various ligands, followed by formation of multicellular aggregates. Following adhesion of one region of the cell to fibronectin-coated beads, the entire surface of the cells became competent to mediate cell-cell aggregation. Aggregates formed in the presence of metabolic inhibitors or signal transduction inhibitors but were reduced in the presence of 8-anilino-1-naphthalene-sulfonic acid (ANS) or Congo Red (CR), perturbants that inhibit protein structural transitions. These perturbants also inhibited aggregation of C. albicans. An increase in ANS fluorescence, which accompanied Als-dependent cellular adhesion, indicated an increase in cell surface hydrophobicity. In addition, C. albicans and Als5p-expressing S. cerevisiae showed an aggregation-induced birefringence indicative of order on the cell surface. The increase in birefringence did not occur in the presence of the aggregation disruptants ANS and CR. These results suggest a model for Als5p-mediated aggregation in which an adhesion-triggered change in the conformation of Als5p propagates around the cell surface, forming ordered aggregation-competent regions.

Substrate concentration‐independent aerobic granulation in sequential aerobic sludge blanket reactor

The development of aerobic granules was studied in four column‐type sequential aerobic sludge blanket reactors fed with different substrate concentrations ranging from 500 to 3000 mg l−1 COD. Results showed that aerobic granules successfully formed in all reactors fed with different substrate concentrations, indicating that the formation of aerobic granules is independent of the substrate concentration. The granule size, roundness, compactness, physical strength, as well as cell surface hydrophobicity and cell polysaccharides contents of the cultivated aerobic granules were investigated. It was shown that aerobic granules formed with different substrate concentrations had similar roundness and compactness. However, the size of aerobic granules slightly increased with an increase in substrate concentration, while granule strength decreased with substrate concentration. It was found that there was a significant increase in cell surface hydrophobicity and cell polysaccharides of the aerobic granules compared to that of seed sludge. The high cell surface hydrophobicity and high cell polysaccharides contents were believed to play an important role in the formation of aerobic granules. However, substrate concentration seems not to be a governing factor for the formation of aerobic granules. The results of this study would be useful for developing aerobic granules‐based bioreactor and for better understanding of the mechanism of aerobic granulation. It was also clearly demonstrated that aerobic granules‐based bioreactor would have great potential in the treatment of high‐strength wastewater.

Variability in Pseudomonas aeruginosa lipopolysaccharide expression during crude oil degradation.

Bacterial utilization of crude oil components, such as the n-alkanes, requires complex cell surface adaptation to allow adherence to oil. To better understand microbial cell surface adaptation to growth on crude oil, the cell surface characteristics of two Pseudomonas aeruginosa strains, U1 and U3, both isolated from the same crude oil-degrading microbial community enriched on Bonny Light crude oil (BLC), were compared. Analysis of growth rates demonstrated an increased lag time for U1 cells compared to U3 cells. Amendment with EDTA inhibited U1 and U3 growth and degradation of the n-alkane component of BLC, suggesting a link between cell surface structure and crude oil degradation. U1 cells demonstrated a smooth-to-rough colony morphology transition when grown on BLC, while U3 cells exhibited rough colony morphology at the outset. Combining high-resolution atomic force microscopy of the cell surface and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extracted lipopolysaccharides (LPS), we demonstrate that isolates grown on BLC have reduced O-antigen expression compared with that of glucose-grown cells. The loss of O-antigen resulted in shorter LPS molecules, increased cell surface hydrophobicity, and increased n-alkane degradation.

Activated carbon surface modifications by adsorption of bacteria and their effect on aqueous lead adsorption

AbstractThe adsorption of Escherichia coli on different activated carbons has been studied. The activated carbon samples used have been characterized, determining their surface area, pore size distribution, elemental analysis, mineral matter analysis and pH of the point of zero charge. The adsorption capacity of these carbons increased with their hydrophobicity and macropore volume. The number of bacteria adsorbed on the demineralized activated carbon in a solution of pH value equal to the iso‐electric point of the carbon was negligible. However, in the presence of cations the proportions of bacterial cells adsorbed were 87.8% (Fe3+), 54.7% (Ca2+) and 24.8% (Mg2+) respectively. This increase in adsorption capacity in the presence of electrolytes has been explained on the basis of both the reduction in electrostatic free energy and the increase in cell surface hydrophobicity due to the metal bound by some compounds of the cell membrane. When the solution pH was intermediate between the pH values of the point of zero charge of the carbon and bacteria the number of bacteria adsorbed increased due to the attractive interactions between the carbon and bacteria. The adsorption of bacteria on activated carbons decreased the porosity and increased the negative charge density of the latter. Depending on the experimental conditions used, the presence of bacteria can enhance the capacity of activated carbons to adsorb lead.© 2001 Society of Chemical Industry

Analysis of functional domains of the Enterococcus faecalis pheromone-induced surface protein aggregation substance.

Pheromone-inducible aggregation substance (AS) proteins of Enterococcus faecalis are essential for high-efficiency conjugation of the sex pheromone plasmids and also serve as virulence factors during host infection. A number of different functions have been attributed to AS in addition to bacterial cell aggregation, including adhesion to host cells, adhesion to fibrin, increased cell surface hydrophobicity, resistance to killing by polymorphonuclear leukocytes and macrophages, and increased vegetation size in an experimental endocarditis model. Relatively little information is available regarding the structure-activity relationship of AS. To identify functional domains, a library of 23 nonpolar 31-amino-acid insertions was constructed in Asc10, the AS encoded by the plasmid pCF10, using the transposons TnlacZ/in and TnphoA/in. Analysis of these insertions revealed a domain necessary for donor-recipient aggregation that extends further into the amino terminus of the protein than previously reported. In addition, insertions in the C terminus of the protein also reduced aggregation. As expected, the ability to aggregate correlates with efficient plasmid transfer. The results also indicated that an increase in cell surface hydrophobicity resulting from AS expression is not sufficient to mediate bacterial aggregation.