Bacterial Extracellular Polymeric Substances Research Articles

Nanoscale Spatiotemporal Dynamics of Microbial Adhesion: Unveiling Stepwise Transitions with Plasmonic Imaging.

Understanding bacterial adhesion at the nanoscale is crucial for elucidating biofilm formation, enhancing biosensor performance, and designing advanced biomaterials. However, the dynamics of the critical transition from reversible to irreversible adhesion has remained elusive due to analytical constraints. Here, we probed this adhesion transition, unveiling nanoscale, step-like bacterial approaches to substrates using a plasmonic imaging technique. This method reveals the discontinuous nature of adhesion, emphasizing the complex interplay between bacterial extracellular polymeric substances (EPS) and substrates. Our findings not only deepen our understanding of bacterial adhesion but also have significant implications for the development of theoretical models for biofilm management. By elucidating these nanoscale step-like adhesion processes, our work provides avenues for the application of nanotechnology in biosensing, biofilm control, and the creation of biomimetic materials.

Interface Engineering of PdPt Ultrafine Ethanol Electro-Oxidation Nanocatalysts by Bacterial Soluble Extracellular Polymeric Substances (s-EPS) to Break through Sabatier Principle.

Unsatisfactory performance of ethanol oxidation reaction (EOR) catalysts hinders the application of direct ethanol fuel cells (DEFCs), while traditional alloy catalysts (like PdPt) is cursed by Sabatier principle due to countable active site types. However, bacterial soluble extracellular polymeric substances (s-EPS) owning abundent functional groups may help breacking through it by contrusting different active sites on PdPt and inducing them to play synergy effect, which is called interface engineering. Using s-EPS to engineer catalysts is more green and consumes lower energy compared to chemical reagents. Herein, PdPt alloy nanoparticles (≈2.1nm) are successfully in situ synthesized by/on s-EPS of Bacillus megaterium, an ex-holotype. Tryptophan residuals are proved as the main reductant. In EOR, PdPt@s-EPS shows higher activity (3.89mAcm-2 ) than Pd@s-EPS, Pt@s-EPS, Pt/C and most reported akin catalysts. Its stability and durability are excellent, too. DFT modelling further demonstrates that, interface engineering by s-EPS breaks through Sabatier principle, by the synergy of diverse sites owning different degrees of d-p orbital hybridization. This work not only makes DEFCs closer to practice, but provides a facile and green strategy to design more catalysts.

The dynamics of bacterial proliferation, viability, and extracellular polymeric substances in oral biofilm development

ObjectivesThis study investigated the relationship between bacterial growth, viability, and extracellular polymeric substances (EPS) formation in biofilms, particularly regarding resistance development. It also examined the impact of chemical factors on the EPS matrix and bacterial proliferation in oral biofilms. MethodsThree multi-species oral biofilms were incubated in anaerobic conditions. Three strains of Enterococcus faecalis were incubated in aerobic conditions. The incubation periods ranged from 0 h to 7 days for short-term biofilms, and from 3 to 90 days for long-term biofilms. Fluorescent labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry were used to track EPS and bacterial growth. Confocal laser scanning microscopy (CLSM) assessed bacterial viability and EPS structure. Biofilms aged 7, 14, and 21 days were treated with 2 % chlorhexidine (CHX) and 1 % sodium hypochlorite (NaOCl) to evaluate their effects on EPS and bacterial proliferation. ResultsShort-term biofilms showed rapid bacterial proliferation and a gradual increase in EPS, maintaining stable viability. In the first two weeks, a significant rise in CFSE indicated growing maturity. From 14 to 90 days, EPS and CFSE levels stabilized. Following treatment, CHX significantly reduced bacterial proliferation, while NaOCl decreased EPS volume. ConclusionsBiofilm development involves a balance between bacterial proliferation and EPS production. The complexity of this process poses challenges in treating biofilm-associated infections, requiring strategies tailored to the biofilm's developmental stage. Clinical significanceFor effective root canal treatment, it is imperative to focus on reducing bacterial proliferation during the early stages of oral infections. In contrast, strategies aimed at minimizing EPS production could be more beneficial for long-term management of these conditions.

Galactosamine and mannosamine are integral parts of bacterial and fungal extracellular polymeric substances.

Extracellular polymeric substances (EPS) are produced by microorganisms and interact to form a complex matrix called biofilm. In soils, EPS are important contributors to the microbial necromass and, thus, to soil organic carbon (SOC). Amino sugars (AS) are used as indicators for microbial necromass in soil, although the origin of galactosamine and mannosamine is largely unknown. However, indications exist that they are part of EPS. In this study, two bacteria and two fungi were grown in starch medium either with or without a quartz matrix to induce EPS production. Each culture was separated in two fractions: one that directly underwent AS extraction (containing AS from both biomass and EPS), and another that first had EPS extracted, followed then by AS determination (exclusively containing AS from EPS). We did not observe a general effect of the quartz matrix neither of microbial type on AS production. The quantified amounts of galactosamine and mannosamine in the EPS fraction represented on average 100% of the total amounts of these two AS quantified in cell cultures, revealing they are integral parts of the biofilm. In contrast, muramic acid and glucosamine were also quantified in the EPS, but with much lower contribution rates to total AS production, of 18% and 33%, respectively, indicating they are not necessarily part of EPS. Our results allow a meaningful ecological interpretation of mannosamine and galactosamine data in the future as indicators of microbial EPS, and also attract interest of future studies to investigate the role of EPS to SOC and its dynamics.

Selenium bio-nanocomposite based on extracellular polymeric substances (EPS): Synthesis, characterization and application in alleviating cadmium toxicity in rice (Oryza sativa L.)

Selenium nanoparticles (SeNPs) have gained significant attention owing to their favorable bioavailability and low toxicity, making them widely applications in the fields of medicine, food and agriculture. In this study, bacterial extracellular polymeric substances (EPS) were used as a novel stabilizer and capping agent to prepare dispersed SeNPs. Results show that EPS-SeNPs presented negative potential (−38 mV), spherical morphologies with average particle size about 100–200 nm and kept stable at room temperature for a long time. X-ray diffraction (XRD) analysis demonstrated that the synthesized nanoparticles were pure amorphous nanoparticles, and X-ray photoelectron spectroscopy (XPS) spectrum showed a spike at 55.6 eV, indicating the presence of zero-valent nano‑selenium. Fourier-transform infrared spectroscopy (FTIR) and three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectroscopy analysis confirmed proteins and polysaccharides in EPS played a crucial role in the synthesis of EPS-SeNPs. Compared to EPS or sodium selenite (Na2SeO3), EPS-SeNPs showed a relatively moderate result in terms of scavenging free radicals in vitro. In contrast, EPS-SeNPs demonstrated lower toxicity to rice seeds than Na2SeO3. Notably, the exogenous application of EPS-SeNPs effectively alleviated the growth inhibition and oxidative damaged caused by cadmium (Cd), and significantly reduced Cd accumulation in rice plants.

Phase separation of extracellular polymeric substances induced by bacteria responsive Nano-Antibiotics for drug-resistant bacterial infection management

Bacterial quorum sensing (QS) and bacterial extracellular polymeric substances (EPS) are the major challenges in treating biofilm chronic infection. Herein, we proposed targeting intervention phase separation of EPS by nano-antibiotics (Ir(ppy)2dmb-PAZ-HA) as an effective strategy for combating bacterial biofilm infection, in which Iridium complex (Ir(ppy)2dmb-CHO) and antibiotic (Pazufloxacin, PAZ) for synergistic antibacterial, and hyaluronic acid (HA) was used to enhance Ir(ppy)2dmb-PAZ release on-demand in biofilm microenvironment. In vitro results showed that Ir(ppy)2dmb-PAZ-HA holds excellent antibacterial activity and antibiofilm capacity. Importantly, Ir(ppy)2dmb-PAZ-HA can be target to regulate phase separation of EPS, and then interfere with biofilm formation, which is a competitive mechanism between Ir(ppy)2dmb-PAZ-HA and Methicillin-resistant Staphylococcus aureus (MRSA) itself. In addition, in vivo results confirmed that Ir(ppy)2dmb-PAZ-HA was able to accelerate biofilm chronic wound healing, and effectively alleviate bacterial acute lung infection. Accordingly, this study provides a new antibiofilm target, and provides a novel idea for the design of small molecular metal antibacterial drugs and antibiotic reuse.

Nanobiopolymer: Potential Applications in Bioremediation of Cadmium Contaminated Water

Nanobiopolymer nanotechnology, bacterial extracellular polymeric substances (EPS), has been used markedly for various water treatments.EPS has polysaccharides, proteins, and lipids, which have functional groups such as amino, sulfhydryl, carboxylic, and phosphate groups that allow binding to metal ions. The accumulation of the metal ions onto the EPS bacterial cell walls is used widely in the bioremediation of cadmium and other metals. This study objectives were extracting EPSs from the bacterium Bacillus subtilis 168 Cd2 and investigating their cadmium adsorption in bead column reactors after being immobilized with calcium alginate. For comparison with the bacterium's role, B. subtilis 168 Cd2 was used before EPS extraction and immobilized with calcium alginate, as well as a comparison with the use of calcium alginate. Cadmium adsorption from solution onto calcium alginate beads with untreated B. subtilis 168 Cd2 and EPS-free B. subtilis 168 Cd2 and under different pH (4.00, 7.00, and 7.50) were studied. The percentages of adsorption onto untreated B. subtilis 168 Cd2 and EPS-free B. subtilis 168 Cd2 were determined and were 89% and 77%, respectively. The adsorption of these were reduced when the pH increased. Scanning electron microscopy (SEM) confirmed the changes in the morphology of the adsorbents, and X-ray fluorescence (XRF) spectrometry analysis recorded the cumulative concentrations of cadmium in the adsorbents after the cadmium adsorption process. EPS in untreated B. subtilis 168 Cd2 significantly enhanced cadmium adsorption percentages. The fewer carboxyl and phosphate groups in EPS-free B. subtilis 168 Cd2 decreased cadmium adsorption. The results obtained in this study are of fundamental significance for applying nano biopolymers nanotechnology environmental biotechnology and bioremediation.

Enhanced disintegration mechanism of surplus activated sludge to improve dewatering by thermally activated persulfate oxidation under mild temperature.

The dewatering treatment is an essential process for the treatment and disposal of surplus activated sludge (SAS), and improving sludge dewatering performance is still a challenge and has become a research hotspot in recent years. The oxidation and disintegration of bacterial cells and extracellular polymeric substances (EPS) by active radicals produced by advanced oxidation processes (AOPs) were extremely promising to achieve deep sludge dewatering. This paper systematically studied the efficiency and mechanism of thermally activated persulfate (TAP) oxidation technology to the improvement of SAS dewatering performance. The results showed that the relative filterability (CST0/CST) was increased 2.52 times with 2.0 mmol/g VSS potassium peroxydisulfate (PDS) at 80 °C in 90 min. Under this condition, the Zeta potential of SAS significantly decreased from - 14.8 to - 1.44 mV, while the average particle size (dp50) decreased from 52.981 to 48.259 μm. Thermal treatment disrupted the sludge structure to release large amounts of EPS including polysaccharides and protein. Meanwhile, the results of three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectra showed that the TAP treatment could expedite the disintegration of sludge, facilitating the decrease of total EPS content and conversion of tightly bound EPS (TB-EPS) to loosely bound EPS (LB-EPS) and soluble EPS (S-EPS). The mechanism of TAP process to improve SAS dewatering performance was revealed, which could contribute to breaking the bottleneck of sludge depth dewatering and provide a theoretical and technical basis for its practical application.

Enhanced membrane fouling by microplastics during nanofiltration of secondary effluent considering secretion, interaction and deposition of extracellular polymeric substances

Microplastic (MP) has been found to influence membrane fouling during microfiltration/ultrafiltration processes in direct and indirect ways by acting as fouling components and changing microbial activities, respectively. However, there is no relevant research about the contribution of MPs to nanofiltration membrane fouling. In this study, for the first time, the impacts of MPs on membrane fouling during the nanofiltration of secondary effluent (SE) were systematically investigated from the perspective of bacterial extracellular polymeric substances (EPS) secretion, their interaction with coexisting pollutants and also deposition. Membrane flux behaviors indicate that MPs simultaneously aggravated the short-term and long-term membrane fouling resistance of nanofiltration by 46 % and 27 %, respectively. ATR-FTIR, XPS and spectrophotometry spectra demonstrate that the deteriorated membrane fouling by MPs directly resulted from the increased accumulation of protein-like, polysaccharides-like and humic-like substances on membranes. EEM spectra further confirmed that MPs preferred to induce serious cake layers, which dominated membrane flux decline but hindered pore fouling. According to CLSM and SEM-EDS mappings, MPs in SE could stimulate microbial activities and then aggravate EPS secretion, after which their interaction with Ca2+ was also enhanced in bulk solution. The cross-linker nets could promote the deposition of other unlinked pollutants on membranes. Besides, MPs could weaken the rejection of certain dissolved organic matters (from 57 % to 52 % on the 50th day of filtration) by aggravating cake-enhanced concentration polarization (CECP), but improved the average removal of inorganic salts from 58 % to 63 % by improving their back diffusion through cake layers. Based on these analyses, the mechanisms of MP-enhanced membrane fouling during the nanofiltration of SE can be thoroughly revealed.

Antibiotics can alter the bacterial extracellular polymeric substances and surface properties affecting the cotransport of bacteria and antibiotics in porous media

Currently, studies on the environmental impact of antibiotics have focused on toxicity and resistance genes, and gaps exist in research on the effects of antibiotics entering the environment on bacterial surface properties and the synergistic transport of antibiotics and bacteria in porous media. To fill the gaps, we investigated the interactions between bacteria and antibiotics in synergistic transport in saturated porous media and the effects of media particle size, flow rate, and ionic concentration on this synergistic transport. This study revealed that although synergistic transport was complex, the mechanism of action was clear. Antibiotics could affect bacterial extracellular polymeric substances (EPS), thus altering their surface hydrophobicity and roughness, thereby affecting bacterial transport. The effects of antibiotics on bacterial transport were dominated by altering bacterial roughness. Antibiotics had a relatively high adsorption on bacteria, so bacterial transport directly affected antibiotic transport. The antibiotic concentrations below a certain threshold increased the bacterial EPS quality, and above the threshold decreased the bacterial EPS quality. This threshold was related to antibiotic toxicity and bacterial type. Bacterial surface hydrophobicity was determined by the combination of proteins and sugars in the EPS, and roughness was positively correlated with the EPS quality.

Combined effects of bacteria and antibiotics on surface properties and transport of nanoplastics in porous media

Currently, research on the individual effects of bacteria and antibiotics on the transport of nanoplastics (NPs) in porous media is in its infancy, while research on their combined effect is absent. It is well known that bacteria and antibiotics also interact with each other, so this synergistic transport of bacteria, antibiotics, and NPs in porous media must be very interesting. For exploring this aspect, we investigated the individual and combined effects of bacteria and antibiotics on the transport of polystyrene NPs (PS-NPs) in saturated porous media. Hydrophobicity, roughness, and the Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energy were measured and calculated. The PS-NPs' transport in porous media was fitted using a mathematical model. Enhanced roughness and size of PS-NPs with increased bacterial concentration dominated and inhibited the PS-NPs' transport in porous media, although the hydrophilicity of PS-NPs and the energy barrier between PS-NPs and porous media were also increased. An increase in antibiotic concentration reduced the energy barrier between PS-NPs and porous media, thereby decreasing the PS-NPs' transport. Combined effects of bacteria and antibiotics on the PS-NPs' transport were complex and distinct from individual effects, but the mechanisms were clear. Roughness and hydrophilicity of PS-NPs and the DLVO interaction energy between PS-NPs and porous media together influenced this process. In the presence of bacteria, antibiotics could alter the bacterial surface roughness by altering bacterial extracellular polymeric substances, and thus alter the PS-NPs' surface roughness, thereby affecting the PS-NPs' transport in porous media. When antibiotics were present, enhanced bacterial concentration increased the PS-NPs' hydrophilicity and the energy barrier between PS-NPs and porous media, thus promoting the PS-NPs' transport. The findings of this study provided a theoretical basis for clarifying the transport of NPs in porous media under complex environments, facilitating a reduction in environmental pollution of NPs.

Enhanced biofilm-mediated degradation of carcinogenic and mutagenic azo dye by novel bacteria isolated from tannery wastewater

Bacterial biofilm and extracellular polymeric substances (EPS) has emerged as an attractive solution for removing dyes from wastewater. Objectives of this study were to identify the biofilm forming bacteria (BFB), contents of the biofilm-EPS, and quantify the effect of novel BFB on biodegradation of Congo red from synthetic wastewater. In this study, 53.3% (16 out of 30) bacterial strains built fragile – robust biofilms in the glass test tubes. Biochemical (Congo red- and Calcofluor binding), Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy results revealed biofilm-EPS consist of curli fimbriae, cellulose, proteins, lipids, nucleic acids and peptidoglycans. Bacterial strains those decolorized > 90% of azo dyes were sequenced. Four novel azo dye-decolorizing biofilm forming bacteria including Bacillus circulans ES6T, Staphylococcus cohnii ES9T, Proteus pennerii ES17T, ES24T, ES25T and ES31T, and Klebsiella quasipneumonae ES22T were identified. Color reduction of Congo red (CR) by ES6T, ES9T, ES17T and ES22T was optimized using the single factor batch experiments. These bacterial strains under optimum conditions (i.e., salt-optimized broth containing 2% sucrose, 80 mg L−1 CR, 1.5% NaCl, at pH 8, 35 ºC, under microaerophilic condition within 72 h of incubation) removed 96.8 – 99.4% CR. Synthesis of total azoreductase (39.6–43.4-fold) and laccase (22.5–33.2-fold) enzymes was induced under optimal conditions. The CR degradation was validated using FTIR and UV-Vis spectral studies. This is the first report on decolorization and degradation of CR by biofilm-EPS producing bacterial strains of B. circulans ES6T, S. cohnii ES9T, P. pennerii ES17T and K. quasipneumonae ES22T.

Rhamnolipid-induced alleviation of bioclogging in Managed Aquifer Recharge (MAR): Interactions with bacteria and porous media

The prevention and treatment of bioclogging is of great significance to the application of Managed Aquifer Recharge (MAR). This study investigated the alleviating effect of biosurfactant rhamnolipid (RL) on bioclogging by laboratory-scale percolation experiments. The results show that the addition of RL greatly reduced bioclogging. Compared with the group without RL, the relative hydraulic conductivity (K′) of the 100 mg/L RL group increased 5 times at the end of the experiment (23 h), while the bacterial cell amount and extracellular polymeric substances (EPS) content on the sand column surface (0–2 cm) decreased by 60.8% and 85.7%, respectively. In addition, the richness and diversity of the microbial communities within the clogging matter decreased after the addition of RL. A variety of bacterial phyla were found, among which Proteobacteria were predominant in all groups. At the genus level, RL reduced the relative abundance of Acinetobacter, Bacillus, Klebsiella, and Pseudomonas. These microbes are known as strong adhesion, large size, and easy to form biofilms, therefore playing a critical role during MAR bioclogging. Moreover, RL changed the surface properties of bacteria and porous media, which results in the increase of electrostatic repulsion and decrease of hydrophobic interaction between them. Therefore, RL mediated the bacteria-porous media interaction to reduce biomass in porous media, thereby alleviating bioclogging. This study implies that RL's addition is an environmentally friendly and effective method to alleviate the bioclogging in MAR.

Microplastic biofilm: An important microniche that may accelerate the spread of antibiotic resistance genes via natural transformation

Microplastic (MP) biofilms provide a specific microniche for microbial life and are a potential hotspot for the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs). Nevertheless, the acquisition of ARGs in MP biofilms via natural transformation mediated by extracellular DNA (eDNA) has been rarely explored. This study demonstrated that MP biofilms promoted the natural transformation of extracellular ARGs at the single-cell and multi-species levels, compared to natural substrate (NS) biofilms and bacterioplankton. The transformation frequency on MP biofilms was up to 1000-fold compare to that on NS. The small MPs and aged MPs enhanced the ARG transformation frequencies up to 77.16-fold and 32.05-fold, respectively, compared with the large MPs and pristine MPs. The transformation frequencies on MP biofilms were significantly positively correlated with the bacterial density and extracellular polymeric substance (EPS) content (P < 0.05). Furthermore, MPs significantly increased the expression of the biofilm formation related genes (motA and pgaA) and DNA uptake related genes (pilX and comA) compared to NS and bacterioplankton. The more transformants colonized on MPs contributed to the enhanced transformation frequencies at the community-wide level. Overall, eDNA-mediated transformation in MP biofilms may be an important path of ARG spread, which was promoted by heterogeneous biofilm.

Reducing the matrix effect in mass spectral imaging of biofilms using flow-cell culture.

The interactions between soil microorganisms and soil minerals play a crucial role in the formation and evolution of minerals and the stability of soil aggregates. Due to the heterogeneity and diversity of the soil environment, the under-standing of the functions of bacterial biofilms in soil minerals at the microscale is limited. A soil mineral-bacterial biofilm system was used as a model in this study, and it was analyzed by time-of-flight secondary ion mass spectrometry (ToF-SIMS) to acquire molecular level information. Static culture in multi-wells and dynamic flow-cell culture in microfluidics of biofilms were investigated. Our results show that more characteristic molecules of biofilms can be observed in SIMS spectra of the flow-cell culture. In contrast, biofilm signature peaks are buried under the mineral components in SIMS spectra in the static culture case. Spectral overlay was used in peak selection prior to performing Principal component analysis (PCA). Comparisons of the PCA results between the static and flow-cell culture show more pronounced molecular features and higher loadings of organic peaks of the dynamic cultured specimens. For example, fatty acids secreted from bacterial biofilm extracellular polymeric substance are likely to be responsible for biofilm dispersal due to mineral treatment up to 48h. Such findings suggest that the use of microfluidic cells to dynamically culture biofilms be a more suitable method for reducing the matrix effect arisen from the growth medium and minerals as a perturbation fac-tor for improved spectral and multivariate analysis of complex mass spectral data in ToF-SIMS. These results show that the interaction mechanism between biofilms and soil minerals at the molecular level can be better studied using the flow-cell culture and advanced mass spectral imaging techniques like ToF-SIMS.

Inhibition of Staphylococcus aureus biofilms by poly-L-aspartic acid nanoparticles loaded with Litsea cubeba essential oil

Staphylococcus aureus (S. aureus) biofilms contamination on various food-contacting surfaces is considered a significant threat in the field of food. Poly-L-aspartic acid (PASP) was proven to damage biofilm by affecting bacterial adhesion, metabolic activity, and extracellular polymeric substances in this study. Especially for eDNA, its generation was reduced by 49.4 %. After treatment with 5 mg/mL of PASP, the number of S. aureus in the biofilm at different growth stages decreased by 1.20–1.68 log CFU/mL. The nanoparticles prepared by PASP and hydroxypropyl trimethyl ammonium chloride chitosan were used to embed LC-EO (EO@PASP/HACCNPs). The results indicated that the particle size of the optimized nanoparticles was 209.84 nm with an encapsulation rate of 70.28 %. Compared to LC-EO alone, EO@PASP/HACCNPs had more significant permeation and dispersion effects on biofilms and possessed long-lasting anti-biofilm activity. For the biofilm grown for 72 h, the population of S. aureus in the EO@PASP/HACCNPs-treated biofilm was additionally reduced by 0.63 log CFU/mL compared with the LC-EO-treated group. EO@PASP/HACCNPs were also applied to different food-contacting materials. The lowest inhibition rate of EO@PASP/HACCNPs on S. aureus biofilm still reached 97.35 %. The sensory properties of the chicken breast were not affected by EO@PASP/HACCNPs.

Bioreduction of hexavalent chromium via Bacillus subtilis SL-44 enhanced by humic acid: An effective strategy for detoxification and immobilization of chromium

As an organic macromolecule, humic acid (HA) has been extensively used as the protectant for bacteria applied in Cr(VI) microbial remediation. However, the effect of the structural properties of HA on the reduction rate of bacteria and the respective contribution of bacteria and HA to soil Cr(VI) management remained uncertain. In this paper, the structural differences between two kinds of humic acid (AL-HA and MA-HA) were explored by means of spectroscopy and electrochemical characterization, and the potential influence of MA-HA on Cr(VI) reduction rate and physiological characteristics of bacteria (Bacillus subtilis, SL-44) also were analyzed. The results showed that the phenolic groups and carboxyl on the surface of HA are firstly complex with Cr(VI) ions, and the fluorescent component with more п-п conjugate structure in HA is the most sensitive species. Compared with single bacteria, the application of SL-44 and MA-HA complex (SL-MA) not only enhanced the reduction of 100 mg/L Cr(VI) to 39.8 % within 72 h and formation rate of intermediate Cr(V), but also reduced the electrochemical impedance. Moreover, the addition of 300 mg/L MA-HA also relieved the Cr(VI) toxicity and decreased the accumulation of glutathione to 94.51 % in bacterial extracellular polymeric substance, furthermore down-regulated the gene expression related to amino acid metabolism and polyhydroxybutyric acid (PHB) hydrolysis in SL-44. Finally, the application of SL-MA also enhanced the stability of chromium in soil and decreased its phytoavailability to 86.09 %, which further reduced chromium enrichment in cabbage organs. These findings provide new insights into Cr(VI) removal, which is also critical for evaluating the application potential of HA for enhancing Cr(VI) bio-reduction.

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.

Assessment of biodegradation kinetics and mass transfer aspects in attached growth bioreactor for effective treatment of brilliant green dye from wastewater

In this study, Bacillus licheniformis immobilized with low-density polyethylene (LDPE) was employed to degrade Brilliant Green (BG) dye from wastewater in a packed bed bioreactor (PBBR). Bacterial growth and extracellular polymeric substance (EPS) secretion were also assessed under different concentrations of BG dye. The impacts of external mass transfer resistance on BG biodegradation were also evaluated at different flow rates (0.3–1.2 L/h). A new mass transfer correlation JD=5.71NRe-0.3 was proposed to study the mass transfer aspects in attached-growth bioreactor. The intermediates, namely 3- dimethylamino phenol, benzoic acid, 1–4 benzenediol, and acetaldehyde were identified during the biodegradation of BG and, subsequently degradation pathway was proposed. Han - Levenspiel kinetics parameters μmax and Ks were found to be 0.185 per day and 115 mg/L, respectively. The new insight into mass transfer and kinetics support the design of efficiently attached growth bioreactor to treat a wide range of pollutants.

Isolation and Staining Reveal the Presence of Extracellular DNA in Marine Gel Particles.

Marine gel particles (MGP) are amorphous hydrogel exudates from bacteria and microalgae that are ubiquitous in the oceans, but their biochemical composition and function are poorly understood. While dynamic ecological interactions between marine microorganisms and MGPs may result in the secretion and mixing of bacterial extracellular polymeric substances (EPS) such as nucleic acids, compositional studies currently are limited to the identification of acidic polysaccharides and proteins in transparent exopolymer particles (TEP) and Coomassie stainable particles (CSP). Previous studies targeted MGPs isolated by filtration. We developed a new way of isolating MGPs from seawater in liquid suspension and applied it to identify extracellular DNA (eDNA) in North Sea surface seawater. Seawater was filtered onto polycarbonate (PC) filters with gentle vacuum filtration, and then the filtered particles were gently resuspended in a smaller volume of sterile seawater. The resulting MGPs ranged in size from 0.4 to 100 µm in diameter. eDNA was detected by fluorescent microscopy using YOYO-1 (for eDNA), with Nile red (targeting cell membranes) as a counterstain. TOTO-3 was also used to stain eDNA, with ConA to localise glycoproteins and SYTO-9 for the live/dead staining of cells. Confocal laser scanning microscopy (CLSM) revealed the presence of proteins and polysaccharides. We found eDNA to be universally associated with MGPs. To further elucidate the role of eDNA, we established a model experimental MGP system using bacterial EPS from Pseudoalteromonas atlantica that also contained eDNA. Our results clearly demonstrate the occurrence of eDNA in MGPs, and should aid furthering our understanding of the micro-scale dynamics and fate of MGPs that underly the large-scale processes of carbon cycling and sedimentation in the ocean.