Bacterial Extracellular Polymers Research Articles

Pseudomonas Stutzeri may alter the environmental fate of polystyrene nanoplastics by trapping them with increasing extracellular polymers

Pseudomonas Stutzeri (P. stutzeri) is a denitrifying bacterium that is essential in biological nitrogen removal. To study the interaction between P. stutzeri and polystyrene nanoplastics (PS-NPs), the effects of PS-NPs posed on P. stutzeri were evaluated in terms of bacterial growth, physiology, denitrification function and extracellular polymers (EPS) secretion. Results of confocal laser scanning microscopy (LCSM), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and flow cytometry confirmed that PS-NPs were trapped by P. stutzeri. Exposure to PS-NPs inhibited bacterial growth and expression of denitrification-related genes, but unaffected the denitrifying enzyme activities. The enhanced secretion of EPS caused PS-NPs and bacterial aggregation. And the enzyme activity of SOD in P. stutzeri was increased while that of CAT was decreased. The results of flow cytometry showed that high concentrations of PS-NPs increased the complexity of P. stutzeri cells. These results reveal that P. stutzeri may be affected after trapping PS-NPs and alter their environmental fate as well. SynopsisThis study contributes to the understanding of the possible effect of P. stutzeri on the distribution of PS-NPs and illustrates the potential impact of PS-NPs on P. stutzeri.

Co-removal and recycling of Ba2+ and Ca2+ in hypersaline wastewater based on the microbially induced carbonate precipitation technique: Overlooked Ba2+ in extracellular and intracellular vaterite

This study investigates the co-precipitation of calcium and barium ions in hypersaline wastewater under the action of Bacillus licheniformis using microbially induced carbonate precipitation (MICP) technology, as well as the bactericidal properties of the biomineralized product vaterite. The changes in carbonic anhydrase activity, pH, carbonate and bicarbonate concentrations in different biomineralization systems were negatively correlated with variations in metal ion concentrations, while the changes in polysaccharides and protein contents in bacterial extracellular polymers were positively correlated with variations in barium concentrations. In the mixed calcium and barium systems, the harvested minerals were vaterite containing barium. The increasing concentrations of calcium promoted the incorporation and adsorption of barium onto vaterite. The presence of barium significantly increased the contents of O-CO, N-CO, and Ba-O in vaterite. Calcium promoted barium precipitation, but barium inhibited calcium precipitation. After being treated by immobilized bacteria, the concentrations of calcium and barium ions decreased from 400 and 274 to 1.72 and 0 mg/L (GB/T15454–2009 and GB8978–1996). Intracellular minerals were also vaterite containing barium. Extracellular vaterite exhibited bactericidal properties. This research presents a promising technique for simultaneously removing and recycling hazardous heavy metals and calcium in hypersaline wastewater.

Mechanistic approach on comparative biosorption of dyes by extracellular polymer of Ochrobactrum pseudintermedium C1 utilizing waste mineral lubricating oil

Dyes are widely used in manufacturing of various consumer products for their ability to impart characteristic color. However, dyes are also one of the most important hazardous pollutants in this present day. Presence of hydroxyl, phosphate, ether, carbonyl and alkyl groups also enable bacterial extracellular polymers to function as biosorbent. In this study, negatively charged biosorbent EP-C1 produced by Gram-negative bacterium Ochrobactrum pseudintermedium C1 utilizing waste mineral lubricating oil has given different decolorization percentages for triphenylmethane cationic Malachite Green and azoic anionic Congo Red at neutral pH. Time wise absorbances from Ultraviolet-Visible Spectrophotometry has estimated maximum biosorption of Malachite Green after 300 min. Biosorption capacities have differed with initial dye concentrations, biosorbent dosages and contact time. Time taken by each dye to reach saturation has depended upon their charged bearing groups of each dye and biosorbent. Kinetic (pseudo first and pseudo second order) and diffusion (intra particle and liquid film) parameters in presence of different dye concentrations and biosorbent dosages were studied for understanding the biosorption mechanism. Morphological differences in chemical structure, chemical functional groups of EP-C1 and EP-C1 loaded dye was studied by Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy and Zeta potential measurements. A compiled overview on comparative mechanistic biosorption of Malachite Green and Congo Red dyes by a bacterial extracellular polymer in miniature laboratory shake flask experiments was done in this study. Newer advents on biological remediation of dyes with mineral lubricating oil as sole carbon source can add a new feather to the already available bioremediation techniques in mitigation of hazardous wastes.

Fixating lead in coal gangue with phosphate using phosphate-dissolving bacteria: Phosphorus dissolving characteristics of bacteria and adsorption mechanism of extracellular polymer

Controlling and preventing lead pollution is currently the focus of environmental remediation. Coal gangue contains large quantities of lead, and its environmental impact cannot be ignored. This study investigated the tolerance of Stenotrophomonas maltophilia (YZ-1 train) to lead ion and its fixation effect on lead in coal gangue. The fixation mechanism of lead ions by using the YZ-1 train was studied with CaHPO4 and Ca3(PO4)2. The tolerance mechanism and fixation characteristics of the three bacterial extracellular polymers and cell components to lead were analyzed. The results show that the YZ-1 train had a strong resistance to lead ions. The amount of lead released from coal gangue can be reduced by up to 91.1% upon treatment with the YZ-1 train, which can dissolve phosphate minerals to form stable hydroxyapatite (Pb5(PO4)3(OH)) and pyromorphite (Pb5(PO4)3Cl) with lead ions. Tryptophan and tyrosine from cellular components and extracellular polymers with loosely and tightly bound proteins are the main participants in the fixation of lead ions. The by-products of soluble microbes affect the fixation of lead ions in soluble extracellular polymers. The carboxylic acids and carboxylates secreted by bacteria are involved in the adsorption and fixation of lead ions.

Bacterial extracellular polymeric substances: Biosynthesis and interaction with environmental pollutants

Extracellular polymeric substances (EPS) are highly hydrated matrices produced by bacteria, containing various polymers such as polysaccharides, proteins, lipids, and DNA. Extracellular polymer concentrations, ions, and functional groups provide physical stability to the EPS. Constituents of EPS form the three-dimensional architecture and help acquire nutrition for the bacteria. Structural and functional diversity of the extracellular polymer depends on the specific glycosyltransferases, polymerase and transporter proteins. These enzymes are encoded by specific genes present in operons such as crd, alg, wca, and gum reported in Agrobacterium, Pseudomonas, Enterobacteriaceae, and Xanthomonas. The operons regulate the biosynthesis of extracellular polymers such as curdlan, alginate, colonic acid, and xanthan, respectively. Various functional groups in the EPS, such as carbonyl, hydroxyl, phosphoryl, and amide, provide the sorption site for interaction with environmental pollutants. Hydrophobic interactions and coordinate bonds mainly dominate the binding of EPS with environmental pollutants. EPS binds, emulsifies, and solubilizes the organic compounds, enhancing the degradation process. EPS binds with heavy metals through complexation, surface adsorption, precipitation, and ion exchange mechanisms. The biodegradability efficiency and nontoxicity properties of EPS make it an excellent biopolymer for decontaminating environmental pollutants. This review summarizes an overview of the biosynthetic mechanisms and interaction of the bacterial extracellular polymer with environmental pollutants. Interaction mechanisms of pollutants with EPS and EPS-mediated bioremediation will help develop removal applications. Moreover, understanding the genes responsible for EPS production, and implementation of new genetic methodology can be helpful for the enhanced biosynthesis of EPS to control pollution by sequestrating more environmental pollutants.

Enhanced removal of tetrabromobisphenol A by Burkholderia cepacian Y17 immobilized on biochar

Biochar-immobilized bacteria have been widely used to remove organic pollutants; however, the enhanced effect of biochar-immobilized bacteria on tetrabromobisphenol A (TBBPA) removal has not been fully investigated and the removal mechanism remains unclear. In this study, a bacterial strain with high TBBPA degradation ability, Burkholderia cepacian Y17, was isolated from an e-waste disassembly area, immobilized with biochar, and used for the removal of TBBPA. Comparisons were performed of the factors affecting the immobilization and TBBPA removal efficiency, including the biochar preparation temperature, immobilization temperature, and pH. The highest 7-day TBBPA removal efficiency by immobilized bacteria was observed with the most suitable biochar preparation temperature (BC500) and an immobilization pH and temperature of 7 and 35 °C, respectively. The TBBPA removal efficiency reached 59.37%, which was increased by 30.23% and 15.88% compared to that of free and inactivated immobilized Y17, respectively. The suitable biochar preparation temperature BC500, immobilization temperature of 35 °C, and neutral pH of 7 increased the bacterial population and extracellular polymer concentration, which facilitated bacterial immobilization on biochar and promoted TBBPA removal. In this case, the high immobilized bacteria concentration (4.62 × 108 cfu∙g−1) and protein and polysaccharide contents (28.43 and 16.16 mg·g−1) contributed to the removal of TBBPA by facilitating TBBPA degradation. The main TBBPA degradation processes by BC500-immobilized Y17 involved debromination, β-scission, demethylation, O-methylation, hydroxylation, and hydroxyl oxidation. This study proposes a method for preparing immobilized bacteria for TBBPA removal and enriches the microbial degradation technology for TBBPA.

Effect of rare earth element Y(III) on short‐term denitrification performance of anaerobic ammonia oxidized granular sludge

AbstractBACKGROUNDThe ammonia‐nitrogen wastewater generated during rare‐earth mining poses a serious hazard to the environment around the mine. This type of wastewater is difficult to treat using conventional biological methods due to its low carbon‐to‐nitrogen ratio. Anaerobic ammonia oxidation (anammox) has become the first choice for treating rare earth mine wastewater. However, the effect of rare earth elements (REEs) in wastewater on anammox bacteria has been insufficiently studied, and this limits the application of this process. Yttrium is a rare earth element present in wastewater, and the study of its effect on anammox can help in the application of this process in rare earth mine wastewater.RESULTSThe effect of Y(III) on anammox granular sludge was studied using batch experiments. Anammox activity decreased with increasing Y(III) concentration, from 64.78 ± 0.28 to 17.15 ± 2.70 mg g−1 VSS day−1. The semi‐inhibitory concentration (IC50) of Y(III) for anammox activity was 30.36 mg L−1. Y (III) exhibited low promotion and high inhibition of anammox bacterial extracellular polymer secretion (EPS). Comparing the kinetic models, the Loung model is better suited for describing the kinetic equation of substrate concentration on anammox.CONCLUSIONThe acute toxicity of Y(III) to anaerobic sludge is influenced by the substrate and Y (III) concentrations. EPS‐based defence strategies play an important role in counteracting the toxicity of Y(III). The Edwards model fits the experimental data well and is suitable for predicting the inhibition of anammox activity by the substrate concentration. © 2022 Society of Chemical Industry (SCI).

Application of a Spiral Symmetric Stream Anaerobic Bioreactor for treating saline heparin sodium pharmaceutical wastewater: Reactor operating characteristics, organics degradation pathway and salt tolerance mechanism

In this study, a Spiral Symmetry Stream Anaerobic Bioreactor (SSSAB) was adopted for treating actual saline heparin sodium pharmaceutical wastewater (HSPW). After adaptation, under the influent COD of 8731 mg/L, OLR of 6.98 kg COD/(m³•d) and salinity of 3.57 wt%, the COD removal reached up to 82%. This value is much higher than the reported for the other reactors at similar salinity. Benzenes are the major organic compounds in HSPW. The main rate-limiting steps are the degradations of phenol and p-cresol. In addition, the degradation pathways of typical benzenes in HSPW were analyzed. After adaptation, the soluble salt content in the granular sludge increased, and the bacterial extracellular polymers (EPS), especially tightly-bound EPS also significantly increased. 16S rRNA analysis revealed that the microbial community in the anaerobic granular sludge (AGS) had become adapted to the HSPW treatment since Mesotoga (12.4%), Anaerophaga (9.0%), Oceanotoga (6.1%) and Aminobacterium (4.1%) increased from previously below 1.0% values. The relative abundance of Methanosarcina in the upper layer of the reactor (68.7%) is significantly higher than that at the bottom (3.8%). This proves the superiority of the SSSAB structure. Finally, a model for salt-tolerant microorganisms is given, which proposes a mechanism for this study and provides reference for other anaerobic biological treatments of high-salt containing wastewater.

Bacterial Extracellular Polymers: A Review

Prokaryotic microbial cells especially bacteria are highly emphases for their exopolysaccharides (EPS) production. EPS are the higher molecular weight natural extracellular compounds observe at the surface of the bacterial cells. Nowadays bacterial EPS represent rapidly emerging as new and industrially important biomaterials because it having tremendous physical and chemical properties with novel functionality. Due to its industrial demand as well as research studies the different extraction processes have been discovered to remove the EPS from the microbial biofilm. The novelties of EPS are also based on the microbial habitat conditions such as higher temperature, lower temperature, acidic, alkaliphilic, saline, etc. Based on its chemical structure they can be homopolysaccharide or heteropolysaccharide. EPSs have a wide range of applications in various industries such as food, textile, pharmaceutical, heavy metal recovery, agriculture, etc. So, this review focus on the understanding of the structure, different extraction processes, biosynthesis and genetic engineering of EPS as well as their desirable biotechnological applications.

Efficient removal of atrazine by iron-modified biochar loaded Acinetobacter lwoffii DNS32

In order to efficiently remove commonly used herbicide atrazine in farmland, an iron-modified biochar (FeMBC) was fabricated via chemical co-precipitation of Fe3+ onto corn stalks biochar. The composites of FeMBC and Acinetobacter lwoffii DNS32 (bFeMBC) effectively accelerated the degradation rate of atrazine (100 mg L−1) in inorganic salt culture solution. TEM，XRD，XPS and FTIR were used to study the basic properties of the Materials. FeMBC promoted the formation of bacterial biofilm, -NH functional group on the surface of bacterial extracellular polymers (EPS) and FeMBC could interact with the aromatic ring of atrazine through Hbonding, which were conducive for microbial capture of atrazine. Meanwhile, the pores (2–10 μm) of FeMBC facilitated the passage of the DNS32 strain and the atrazine molecule, which contributed to the efficient capture and degradation of atrazine by DNS32 strain. BFeMBC amendment helped to maintain the bacterial diversity in the atrazine contaminated soil. The increase of rare bacteria (relative abundance of 0.01%–0.05%) richness plays a certain role in stabilizing nutrient cycling, thereby promoting microbial nutrient utilization activities and has the function of pollutant degradation. This may contribute to the digestion of atrazine and its intermediate metabolites，reducing the stress of microbial in atrazine contaminated soil. bFeMBC amendment may be a promising in situ remediation technique for soil atrazine contamination.

Time Evolution of Bacterial Extracellular Polymer Composition Studied with a Versatile Infrared Transmission Array Scanner

The chemical composition of extracellular polymeric substances (EPSs) secreted by Escherichia coli K12 in solution was monitored as a function of time over various stages of bacterial growth. We ha...

Contaminants removal by bentonite amended slow sand filter

Earlier studies have indicated that variability in size, surface texture and charge greatly influence the contaminant removal process in granular media. Based on surface characteristics of montmorillonite, it is anticipated that small addition of this clay would increase adhesion sites for bacterial growth and extracellular polymer production in the slow sand filter and thereby enhance its contaminant removal ability. Experiments were performed by permeating groundwater contaminated with pathogens (total coliform and E. Coli) and inorganic contaminants through the bentonite amended slow sand filter (BASSF). Surprisingly, the BASSF retained inorganic contaminants besides pathogens. Water-leach tests (pH of water leachate ranged from 2 to 9) with spent BASSF specimen indicated that the inorganic contaminants are irreversibly adsorbed to a large extent. It is considered that the combined effects of enhanced-organic matter mediated adhesion sites and increased hydraulic retention time enables the BASSF specimen to retain inorganic contaminants. It is envisaged that BASSF filters could find use in treating contaminated groundwater for potable needs at household and community level.

Dynamics of dewaterability and bacterial populations in activated sludge

Relationships of bacterial populations and extracellular polymer substances (EPS) to dewaterability of activated sludge were studied on three laboratory-scale activated sludge reactors fed with synthetic wastewater. Dewaterability of activated sludge was evaluated by a novel method developed by the authors, in which small amount of sludge was centrifugally dewatered, and its water content was measured. Bacterial populations during the reactor operation were analyzed by the polymerase chain reaction/terminal-restriction fragment length polymorphism (PCR/T-RFLP) targeted at a partial 16S rRNA gene. Extracellular polymeric substances (EPS) were extracted using cation exchange resin (CER), and polysaccharides and total protein in EPS were determined. Some of the dominant terminal-restriction fragments (T-RFs) were observed to have significant relationships with dewaterability of sludge, and it was suggested that bacterial species corresponding to those peaks significantly affected dewaterability. On the other hand, significant relationships were not found between EPS concentration and dewaterability of sludge.

Variations of both bacterial community and extracellular polymers: The inducements of increase of cell hydrophobicity from biofloc to aerobic granule sludge

To investigate the inducements of increase of cell hydrophobicity from aerobic biofloc (ABF) and granular sludge (AGS), in this study, as the first time the hydrophilic and hydrophobic bacterial communities were analyzed independently. Meanwhile, the effect of extracellular polymers (EPS) on the cell hydrophobicity is also studied. Few Bacteroidetes were detected (1.35% in ABF and 3.84% in AGS) in hydrophilic bacteria, whereas they are abundant in the hydrophobic cells (47.8% and 43% for ABF and AGS, respectively). The main species of Bacteroidetes changed from class Sphingobacteria to Flavobacteria in AGS. On the other hand, EPS is directly responsible to cell hydrophobicity. For AGS, cell hydrophobicity was sharply decreased after EPS extraction. Both quantity and property of the extracellular protein are related to hydrophobicity. Our results showed the variation of cell hydrophobicity was resulted from variations of both bacterial population and EPS.

Imaging Hydrated Microbial Extracellular Polymers: Comparative Analysis by Electron Microscopy

Microbe-mineral and -metal interactions represent a major intersection between the biosphere and geosphere but require high-resolution imaging and analytical tools for investigation of microscale associations. Electron microscopy has been used extensively for geomicrobial investigations, and although used bona fide, the traditional methods of sample preparation do not preserve the native morphology of microbiological components, especially extracellular polymers. Herein, we present a direct comparative analysis of microbial interactions by conventional electron microscopy approaches with imaging at room temperature and a suite of cryogenic electron microscopy methods providing imaging in the close-to-natural hydrated state. In situ, we observed an irreversible transformation of the hydrated bacterial extracellular polymers during the traditional dehydration-based sample preparation that resulted in their collapse into filamentous structures. Dehydration-induced polymer collapse can lead to inaccurate spatial relationships and hence could subsequently affect conclusions regarding the nature of interactions between microbial extracellular polymers and their environment.

Escherichia coli transport in porous media: Influence of cell strain, solution chemistry, and temperature

Packed bed column and complementary characterization experiments were carried out with two Escherichia coli strains (D21g and XL1-Blue) under a range of ionic strength (IS) and valence (KCl, CaCl 2, and artificial groundwater) to determine the role of bacterial strain and solution chemistry on cell adhesion. Increasing IS and valence had a marked effect on the electrokinetic and surface properties of bacteria and quartz grains; hence resulting in a greater rate of deposition. Distinct deposition trends were observed for the two cell strains, with greater retention observed for D21g versus XL1-Blue across the range of IS. Selected transport and characterization experiments were also conducted with the D21g cells, finding deposition also increasing with IS and valence. In the presence of Ca 2+ bacterial deposition behavior deviated from anticipated trends and it is concluded from additional analysis that Ca 2+ ions influence bacterial surface charge, hydrophobicity, and extracellular polymers. Further transport experiments were conducted with the D21g cells and colloids to establish the role of temperature (4, 10 and 25 °C). Results suggested that a combination of specific and non-specific interactions occurring between the cells and quartz determines the extent of deposition, rather than transport from the bulk to the collector surface.

Bacterial extracellular polymeric substances (EPS) mediate CaCO 3 morphology and polymorphism

The effect of bacterial extracellular polymers (EPS) on calcium carbonate morphology and polymorphism was investigated by means of free-drift mineralisation experiments, conducted using cells of the EPS-producing thermotolerant Bacillus licheniformis S-86. The cells either had the EPS layer intact (native cells) or had the EPS layer removed (EPS-free cells). Experiments were also undertaken in the absence of cells, but with the addition of the extracted EPS solution. It was found that the abiotic control and EPS-free cell experiments contained vaterite and calcite when sampled after 12 h and 48 h, whereas the native cell and EPS-solution experiments contained only calcite. These results suggest that the presence of EPS inhibits vaterite precipitation. It is proposed that dissolved organic carbon (DOC) released from the EPS complexes Ca 2+ ions in solution, reducing the calcium carbonate saturation and favouring calcite precipitation over vaterite. After 7 days, all experiments contained only calcite. However, distinct morphological differences between the calcite crystals precipitated in the four experiments were preserved after a period of 7 days, suggesting that crystal morphology may indicate the biological conditions under which calcite precipitated.

Predicting the rate and extent of cadmium and copper desorption from soils in the presence of bacterial extracellular polymer

The movement of cationic transition metals through the subsurface is strongly retarded by sorption to the porous media. However, dissolved organic ligands can compete with soil surfaces by providing binding sites for metals in solution. An extracellular polymer produced by a bacterium isolated from soil was used in this study to observe and model the influence of a naturally occurring ligand on the release of adsorbed metals from two test soils. Experimental results show that the presence of dissolved extracellular polymer enhanced the rate and extent of desorptive release of soil-bound cadmium and copper. A kinetic model that uses a gamma distribution of rate constants to account for the physical and chemical heterogeneity of the soil matrix was employed to describe the release of cadmium and copper in batch experiments. Model parameters describing soil, metal and extracellular polymer interactions were obtained through separate experiments. With these parameters the model successfully predicted the influence of dissolved polymer on the rate and extent of release of cadmium and copper from soil in independent batch experiments. These results suggest that the presence of natural metal-binding ligands such as bacterial extracellular polymers can act to increase the driving force for desorption by lowering the aqueous concentration of free unbound metals in solution.

Predicting the rate and extent of cadmium and copper desorption from soils in the presence of bacterial extracellular polymer

The movement of cationic transition metals through the subsurface is strongly retarded by sorption to the porous media. However, dissolved organic ligands can compete with soil surfaces by providing binding sites for metals in solution. An extracellular polymer produced by a bacterium isolated from soil was used in this study to observe and model the influence of a naturally occurring ligand on the release of adsorbed metals from two test soils. Experimental results show that the presence of dissolved extracellular polymer enhanced the rate and extent of desorptive release of soil-bound cadmium and copper. A kinetic model that uses a gamma distribution of rate constants to account for the physical and chemical heterogeneity of the soil matrix was employed to describe the release of cadmium and copper in batch experiments. Model parameters describing soil, metal and extracellular polymer interactions were obtained through separate experiments. With these parameters the model successfully predicted the influence of dissolved polymer on the rate and extent of release of cadmium and copper from soil in independent batch experiments. These results suggest that the presence of natural metal-binding ligands such as bacterial extracellular polymers can act to increase the driving force for desorption by lowering the aqueous concentration of free unbound metals in solution.

Mobilization of adsorbed copper and lead from naturally aged soil by bacterial extracellular polymers

Sorption of pollutants is a dominant phase transfer process affecting the fate and transport of metals through the subsurface. The movement of contaminants is retarded by sorption to the stationary subsurface porous media and can seriously hinder remediation efforts. Research has shown that the binding of adsorbed metals becomes more pronounced the longer the contaminant is in the subsurface and the release rates of aged metal contaminants have not received the research attention given to freshly added metals in laboratory studies. Metal release rates are also influenced by the presence of dissolved ligands that compete with mineral soil surfaces by providing binding sites. Dissolved organic matter such as bacterial extracellular polymers are common in natural soil solutions and the metal binding properties of bacterial polymers are well established. Therefore, binding of metals to dissolved biopolymers may result in mobilization of an adsorbed metal. This is important for cases where the metals are assumed to be relatively immobile such as in the case of land applied biosolids. In addition, naturally occurring adherent bacteria commonly produce extracellular polymers and thus may modify the bioavailability of meal contaminants at the point of their attachment. In this study samples from three sites, one a land applied sludge test site, were used to investigate the ability of bacterial extracellular polymers to release metals from soils with long-term exposures. The presence of ≅200 mg/L bacterial extracellular polymer was found to increase the short-term (less than 350 h) release of Cu and Pb by a factor of 2–4-fold.