Extracellular Polymeric Substances Components Research Articles

Quantitative image analysis for assessing extracellular polymeric substances in activated sludge under atrazine exposure

Extracellular polymeric substances (EPS) play a vital role in biological wastewater treatment systems. This study investigates the impact of herbicide atrazine (ATZ) on the overall performance, EPS yield, composition, and sludge morphology in an activated sludge (AS) system operated in a sequencing batch reactor (SBR). Since conventional methods for analyzing EPS are time-consuming and releases residues, a new approach was developed in this work to evaluate the EPS fractions and components, based on the morphological characterization of the biomass using quantitative image analysis (QIA) technique coupled with multivariate statistics.Results showed that exposure to ATZ inhibit biomass activity in terms of organic matter (COD) and nitrogen removal. Moreover, both tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) increased under ATZ, indicating that microorganisms release EPS as a defense mechanism against environmental changes. The PN/PS ratio also increases, indicating likely increased hydrophobicity in ATZ phases. Furthermore, ATZ phases exhibit a predominance of large aggregates compared to intermediate and small ones, indicating a change in aggregate morphological structure associated with EPS production. The new approach using QIA coupled with partial least squares (PLS) modeling provides accurate predictions of EPS content. The increase in TB-EPS is closely related to the rise of large aggregates in phases exposed to higher ATZ concentrations. The PLS models demonstrate high accuracy for EPS prediction (coefficients of determination, R2 above 0.86), showcasing the feasibility of using QIA for EPS assessment in AS systems. This approach offers significant potential for regular process monitoring and management, providing a more environmentally friendly methodology by eliminating the need for chemical usage.

Molecular spectra and docking simulations investigated the binding mechanisms of tetracycline onto E. coli extracellular polymeric substances

Extracellular polymeric substances (EPS), which were an important fraction of natural organic matter (NOM), played an important role in various environmental processes. However, the heterogeneity, complexity, and dynamics of EPS make their interactions with antibiotics elusive. Using advanced multispectral technology, this study examined how EPS interacts with different concentrations of tetracycline (TC) in the soil system. Our results demonstrated that protein-like (C1), fulvic-like (C2), and humic-like (C3) fractions were identified from EPS. Two-dimensional synchronous correlation spectroscopy (2D-SF-COS) indicated that the protein-like fraction gave faster responses than the fulvic-like fraction during the TC binding process. The sequence of structural changes in EPS due to TC binding was revealed by two-dimensional Fourier Transformation Infrared correlation spectroscopy (2D-FTIR-COS) as follows: 1550 > 1660 > 1395 > 1240 > 1087 cm−1. It is noteworthy that the sensitivity of the amide group to TC has been preserved, with its intensity gradually increasing to become the primary binding site for TC. The integration of hetero-2DCOS maps with moving window 2D correlation spectroscopy (MW2DCOS) provided a unique insight into understanding the correlation between EPS fractions and functional groups during the TC binding process. Moreover, molecular docking (MD) discovered that the extracellular proteins would provide plenty of binding sites with TC through salt bridges, hydrogen bonds, and π-π base-stacking forces. With these results, systematic investigations of the dynamic changes in EPS components under different concentrations of antibiotic exposure demonstrated the advanced capabilities of multispectral technology in examining intricate interactions with EPS in the soil environment.

Enhancing biohydrogen production from xylose through natural FeS2 ore: Mechanistic insights

In this study, we investigated the advantages of utilizing natural FeS2 ore in the context of dark fermentative hydrogen production within a fermentation system employing heat-treated anaerobic granular sludge with xylose as the carbon source. The results demonstrated a significant improvement in both hydrogen production and the maximum rate, with increases of 2.58 and 4.2 times, respectively. Moreover, the presence of FeS2 ore led to a reduction in lag time by more than 2–3 h. The enhanced biohydrogen production performance was attributed to factors such as the intracellular NADH/NAD+ ratio, redox-active components of extracellular polymeric substances, secreted flavins, as well as the presence of hydrogenase and nitrogenase. Furthermore, the FeS2 ore served as a direct electron donor and acceptor during biohydrogen production. This study shed light on the underlying mechanisms contributing to the improved performance of biohydrogen production from xylose during dark fermentation through the supplementation of natural FeS2 ore.

Redox mediator chlorophyll accelerates low-temperature biological denitrification with responses of extracellular polymers and changes in microbial community composition

Low temperatures limit the denitrification wastewater in activated sludge systems, but this can be mitigated by addition of redox mediators (RMs). Here, the effects of chlorophyll (Chl), 1,2-naphthoquinone-4-sulfonic acid (NQS), humic acid (HA), and riboflavin (RF), each tested at three concentrations, were compared for denitrification performance at low temperature, by monitoring the produced extracellular polymeric substances (EPS), and characterizing microbial communities and their metabolic potential. Chl increased the denitrification rate most, namely 4.12-fold compared to the control, followed by NQS (2.62-fold increase) and HA (1.35-fold increase), but RF had an inhibitory effect. Chl promoted the secretion of tryptophan-like and tyrosine-like proteins in the EPS and aided the conversion of protein from tightly bound EPS into loosely bound EPS, which improved the material transfer efficiency. NQS, HA, and RF also altered the EPS components. The four RMs affected the microbial community structure, whereby both conditionally abundant taxa (CAT) and conditionally rare or abundant taxa (CRAT) were key taxa. Among them, CRAT members interacted most with the other taxa. Chl promoted Flavobacterium enrichment in low-temperature activated sludge systems. In addition, Chl promoted the abundance of nitrate reduction genes narGHI and napAB and of nitrite reduction genes nirKS, norBC, and nosZ. Moreover, Chl increased abundance of genes involved in acetate metabolism and in the TCA cycle, thereby improving carbon source utilization. This study increases our understanding of the enhancement of low-temperature activated sludge by RMs, and demonstrates positive effects, in particular by Chl.

Mechanistic insight into the aggregation ability of anammox microorganisms: Roles of polarity, composition and molecular structure of extracellular polymeric substances

The chemical characteristics of extracellular polymeric substances (EPS) of anammox bacteria (AnAOB) play a crucial role in the rapid enrichment of AnAOB and the stable operation of wastewater anammox processes. To clarify the influential mechanisms of sludge EPS on AnAOB aggregation, multiple parameters, including the polarity distribution, composition, and molecular structure of EPS, were selected, and their quantitative relationship with AnAOB aggregation was analyzed. Compared to typical anaerobic sludge (anaerobic floc and granular sludge), the anammox sludge EPS exhibited higher levels of tryptophan-like substances (44.82–56.52 % vs. 2.57–39.81 %), polysaccharides (40.02–53.49 mg/g VSS vs. 30.22–41.69 mg/g VSS), and protein structural units including α-helices (20.70–23.98 % vs. 16.48–19.32 %), β-sheets (37.43–42.98 % vs. 25.78–36.72 %), and protonated nitrogen (Npr) (0.065–0.122 vs. 0.017–0.061). In contrast, it had lower contents of β-turns (20.95–27.39 % vs. 28.17–39.04 %). These biopolymers were found to originate from different genera of AnAOB. Specifically, the α-helix-rich proteins were mainly derived from Candidatus Kuenenia, whereas the extracellular proteins related to tryptophan and Npr were closely associated with Candidatus Brocadia. Critically, these EPS components could drive anammox aggregation through interactions. Substantial amounts of tryptophan-like substances facilitated the formation of β-sheet structures and the exposure of internal hydrophobic clusters, which benefited the anammox aggregation. Meanwhile, extracellular proteins with high Npr content played a pivotal role in the formation of mixed protein-polysaccharide gel networks with the electronegative regions of polysaccharides, which could be regarded as the key component in the maintenance of anammox sludge stability. These findings provide a comprehensive understanding of the multifaceted roles of EPS in driving anammox aggregation and offer valuable insights into the development of EPS regulation strategies aimed at optimizing the anammox process.

Spectroscopic and Microscopic Characterization of Microbial Biofouling on Aircraft Fuel Tanks.

Avoiding microbial contamination and biofilm formation on the surfaces of aircraft fuel tanks is a major challenge in the aviation industry. The inevitable presence of water in fuel systems and nutrients provided by the fuel makes an ideal environment for bacteria, fungi, and yeast to grow. Understanding how microbes grow on different fuel tank materials is the first step to control biofilm formation in aviation fuel systems. In this study, biofilms of Pseudomonas putida, a model Gram-negative bacterium previously found in aircraft fuel tanks, were characterized on aluminum 7075-T6 surfaces, which is an alloy used by the aviation industry due to favorable properties including high strength and fatigue resistance. Scanning electron microscopy (SEM) coupled with energy-dispersive X-ray (EDX) showed that extracellular polymeric substances (EPS) produced by P. putida were important components of biofilms with a likely role in biofilm stability and adhesion to the surfaces. EDX analysis showed that the proportion of phosphorus with respect to nitrogen is higher in the EPS than in the bacterial cells. Additionally, different morphologies in biofilm formation were observed in the fuel phase compared to the water phase. Micro-Fourier transform infrared spectroscopy (micro-FTIR) analysis suggested that phosphoryl and carboxyl functional groups are fundamental for the irreversible attachment between the EPS of bacteria and the aluminum surface, by the formation of hydrogen bonds and inner-sphere complexes between the macromolecules and the aluminum surface. Based on the hypothesis that nucleic acids (particularly DNA) are an important component of EPS in P. putida biofilms, the impact of degrading extracellular DNA was tested. Treatment with the enzyme DNase I affected both water and fuel phase biofilms─with the cell structure disrupted in the aqueous phase, but cells remained attached to the aluminum coupons.

In-situ characterization of dynamic Acidithiobacillus ferrooxidans-pyrite interaction via QCM-D-based method: Cell adhesion and biofilm formation in responding to surface sulfur speciation

Microbe-sulfide mineral interactions (MSMI), often manifested as biofilm formation, are ubiquitous in natural and engineered environments. Such interactions were much dependent upon the evolution of chemical species of mineral surface and microbial extracellular polymeric substances (EPS), and in-situ characterization of which have been a demanding and crucial task. In this study, quartz crystal microbalance with dissipation (QCM-D) was used to real time characterize the adhesion of Acidithiobacillus ferrooxidans onto the pyrite surface after introducing L-cysteine (an important component of EPS). And by combining with monitoring the EPS composition in biofilm by confocal laser scanning microscopy (CLSM) and the surface sulfur species by SEM-EDS and X-ray-photoelectron spectroscopy (XPS) as well as the changes in pH, ORP, [Fe2+] and [Fe3+] in solution during A. ferrooxidans-pyrite interaction, the results show that L-cysteine can lead to the Cys-S-S2Fe complex formation on pyrite surface, and under the dosage of 0.02 to 0.1 g/L of L-cysteine, the Cys-S-S2Fe complex promoted the cell adhesion and biofilm formation of A. ferrooxidans with secretion of more EPS proteins but less EPS glycoconjugates, resulting in the promotion of sulfur biooxidation with inhibition of iron biooxidation, corresponding to no obvious formation of jarosites. The higher dosage of L-cysteine (e.g., 0.2 g/L) inhibited biofilm formation, reduced iron/sulfur oxidation. These results deepen the understanding of the roles of L-cysteine on MSMI. This study demonstrates the QCM-D-based method can be widely used for in situ characterization of MSMI, and opens an alternative pathway for revealing the micro-mechanism of MSMI correlated to the macroscopic effects.

Simultaneous change of microworld and biofilm formation in constructed wetlands filled with biochar

As the regulator of constructed wetlands (CWs), biochar is often used to enhance pollutant removal and reduce greenhouse gas emission. Biochar is proved to have certain effects on microbial populations, but its effect on the aggregation of microbial flocs and the formation of biofilms in the CWs has not been thoroughly investigated. Therefore, the above topics were studied in this paper by adding a certain proportion of biochar in aerated subsurface flow constructed wetlands. The results indicated that after adding biochar in the CWs, pollutant removal was enhanced and the removal rate of NH4+-N was increased from 80.76% to 99.43%. The proportion of hydrophobic components in extracellular polymeric substances (EPS) was reduced by adding biochar from 0.0044 to 0.0038, and the affinity of EPS on CH3-SAM was reduced from 5.736 L/g to 2.496 L/g. The weakened hydrophobic and the reduced affinity of EPS caused the initial attachment of microorganisms to be inhibited. The relative abundance of Chloroflexi was decreased after adding biochar, reducing the dense structural skeleton of biofilm aggregates. Correspondingly, the abundance of Bacteroidetes was increased, promoting EPS degradation. Biochar addition helped to increase the proportion of catalytic active proteins in extracellular proteins and decrease the proportion of binding active proteins, hindering the combination of extracellular proteins and macromolecules to form microbial aggregates. Additionally, the proportions of three extracellular protein structures promoting microbial aggregation, including aggregated chain, β-sheet, and 3-turn helix, were decreased to 23.83%, 38.37% and 7.76%, respectively, while the proportions of random coil and antiparallel β-sheet that inhibited microbial aggregation were increased to 14.11% and 8.11%, respectively. An interesting conclusion from the experimental results is that biochar not only can enhance pollutants removal, but also has the potential of alleviating biological clogging in CWs, which is of great significance to realize the sustainable operation and improve the life cycle of CWs.

Roles of extracellular polymeric substances in the adsorption and removal of norfloxacin during hydrothermal treatment of sewage sludge

Hydrothermal treatment (HT) is promising to remove antimicrobials from sewage sludge (SS); however, the mechanism of antimicrobial degradation during the HT of SS is not fully understood. In this study, the roles of extracellular polymeric substances (EPS) in the removal and transformation of norfloxacin (NOR) during the HT of SS at temperatures of 100 and 160 °C were investigated. The results indicated that the degradation of NOR increased with increasing HT temperature, with maximum NOR removal (52%) achieved at 160 °C. Furthermore, the NOR in sludge showed higher degradation efficiencies than the control as HT temperature was higher than 120 °C. Evident promotion effects of bound-EPS (B-EPS) in sludge were observed on the NOR degradation as HT temperature was higher than 120 °C, leading to the mineralization and deamination of protein-like components in EPS during HT. Beside, the adsorption capacity of NOR during the HT of SS decreased at temperatures higher than 120 °C. The evolution of the spatial structure of B-EPS was predominantly responsible for the adsorption of antimicrobials, a spontaneous process driven mainly by hydrophilic interactions. With the hydrothermal conversion of B-EPS, the electron transfer, and reactive species (3EPS* and ·OH) derived from B-EPS could facilitate the degradation of NOR. In particular, hydrogen bonds between B-EPS and NOR increased the apparent yield of ·OH and accelerated the decarboxylation of NOR during HT at temperatures higher than 120 °C. A toxicity evaluation suggested that HT for NOR degradation could attenuate toxicity, whereas deep oxidation or mineralization would be needed to promote ecosystem safety. These findings provide new insights into the hydrothermal activation of EPS and the interrelated hydrothermal fate of antimicrobials and other toxic pollutants in sludge.

Insights into the EPS production and distribution of planktonic and attached Sulfobacillus thermosulfidooxidans cells during bioleaching

Bacterial leaching can effectively promote the bioleaching of chalcopyrite by secreting extracellular polymeric substances (EPS). In this study, the production and distribution of EPS on the surface of planktonic and attached cells of Sulfobacillus thermosulfidooxidans (S. thermosulfidooxidans) during the process of chalcopyrite concentrate bioleaching were observed in situ using Field emission scanning electron microscope (FE-SEM) and Confocal laser scanning microscope (CLSM). The TOC content and key components of EPS were extracted during the chalcopyrite bioleaching process. Moreover, the main functional groups of extracellular polymorphs were identified through Fourier transmission-infrared spectroscopy (FT-IR). The results showed a similar trend in the evolution of EPS content in planktonic and attached cells. However, the EPS content secreted by the cells attached was more than that of planktonic cells during the entire process of bioleaching. Furthermore, the results of FE-SEM indicated that the EPS secreted by the attached cells is thicker and stickier, with better adsorption performance. The FT-IR spectrum analysis revealed that the O–H, C–H, and CO bonds are the main functional groups of EPS. This work further clarifies the role of EPS in the bioleaching process.

Analysis of the Effects of Surfactants on Extracellular Polymeric Substances

Reservoirs after chemical flooding usually have residual chemicals, which can affect the driving effect of subsequent microbial drives. Among them, the effect of surfactants on the metabolites of oil-recovering bacteria is the most obvious. Therefore, this paper investigates the influence mechanism of sodium dodecyl sulfate (SDS) on the nature and structure of Extracellular Polymeric Substances (EPS) produced by metabolism of Enterobacter cloacae, through a variety of characterization to analysis the components and structure of EPS under SDS stress. The results showed that Enterobacter cloacae was identified as a glycolipid-producing strain, the main components of EPS were polysaccharides and proteins. The polysaccharide composition (%: w/w) was glucosamine, 37.2; glucose, 31.5; rhamnose, 26.3; xylose, 1.7; and unidentified sugar, 3.3; and the main component of proteins was polyglutamic acid. EPS under the stress of SDS showed an increase in the content of functional groups such as -C=O and -COOH and an increase in the cellular particle size, and production of EPS increased by 10.69 × 103 mg/L when the SDS concentration was 2.5 × 102 mg/L; 3D-EEM results showed that the components of all three types of EPS The 3D-EEM results showed that all three types of EPS fractions contained tryptophan and protein-like substances, humic acid-like substances were only distributed in the solubilized extracellular polymers (SL-EPS), and aromatic proteins were only present in the loosely bound type (LB-EPS) and tightly bound type (TB-EPS). In addition, the peaks representing humic-like substances showed a blue shift, indicating that SDS had the greatest effect on SL-EPS. This study provides a guidance for refining the mechanism of strain EPS response to reservoir residual surfactant SDS, and provides a more comprehensive and in-depth understanding of surfactant-protein interactions.

Extracellular DNA enhances biofilm integrity and mechanical properties of mucoid Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the most common biofilm-forming pathogens responsible for lung infections of individuals with cystic fibrosis (CF). P. aeruginosa becomes tolerant to antimicrobials in the biofilm state and is difficult to treat. Production of extracellular polymeric substances (EPS), such as alginate and extracellular DNA (eDNA), can allow adherence to abiotic and biotic surfaces, antimicrobial evasion, and resilience to environmental pressures. Alginate-producing mucoid variants of P. aeruginosa are frequently isolated from CF airway samples and are associated with worsening patient outcomes. While eDNA is a major structural component of nonmucoid P. aeruginosa biofilms, the potential role of eDNA in mucoid biofilms is unclear. Here, we investigate how eDNA contributes to clinical mucoid biofilm physiology and integrity. We predicted that eDNA plays a structural and mechanical role in mucoid biofilms. To test this, we quantified biofilm eDNA in mucoid biofilms and used microscopy and rheology to visualize eDNA and detect changes in biofilm structure and mechanics upon DNaseI treatment. We showed that biofilm eDNA abundance is diverse across clinical mucoid strains and observed a temporal increase in foci of eDNA within intact mucoid biofilms. Increased cell dispersal and reduced biomass were also observed following DNaseI treatment of mucoid biofilms. Degradation of eDNA also impacted the mechanical integrity of mucoid biofilms by increasing the stiffness and decreasing the cohesion of the biofilm. These findings advance our understanding of clinical mucoid P. aeruginosa biofilms and facilitate the development of new approaches to target biofilms by exploiting the functions of EPS components. IMPORTANCE Understanding the role of eDNA in mucoid Pseudomonas aeruginosa biofilms will lead to therapeutic strategies that combat the biophysical and structural function of EPS for the eradication of bacteria in mucoid biofilms during chronic infections. This knowledge can be used to further identify unknown matrix component interactions within pathogenic biofilm-forming clinical isolates.

Extracellular polymeric substances and microbial community shift during the start-up of a single-stage partial nitritation/anammox process.

A sequencing batch reactor (SBR) was operated to investigate variations of extracellular polymeric substances (EPS) and microbial community during the start-up of the single-stage partial nitritation/anammox (SPN/A) process at intermittent aeration mode. The SPN/A system was successfully started on day 34, and the nitrogen removal efficiency and total nitrogen loading rate were 82.29% and 0.31 kg N/(m3 ·d), respectively. Furthermore, the relationship between the protein secondary structures and microbial aggregation was strongly related. The α-helix/ (β-sheet + random coil) ratios increased obviously from 0.20 ± 0.03 to 0.23 ± 0.01, with the sludge aggregation mean size increased from 56 to 107 μm during the start-up of SPN/A. During the start-up of SPN/A, Candidatus Kuenenia was the primary anammox bacteria, whereas Nitrospira was the main functional bacteria of nitrite-oxidizing bacteria. Correlation between the microbial community and EPS components was performed. The EPS and microbial community played important roles in keeping stable nitrogen removal and the formation of sludge granules.

Metagenomics and proteomics profiling of extracellular polymeric substances from municipal waste sludge and their application for soil and water bioremediation

This study assessed the components of anaerobically digested sludge, activated sludge, and microbial and extracellular polymeric substance (EPS) enzymes to identify the mechanisms underlying nitrogen removal and soil regeneration. 16S rRNA gene amplicon-based sequencing was used to determine the microbial community composition and the related National Center for Biotechnology Information (NCBI) protein database was used to construct a conventional library from the observed community. EPS components were identified using gel-free proteomic (Liquid Chromatography with tandem mass spectrometry-LC/MS/MS) methods. Alginate-like EPS from aerobically activated sludge have strong potential for soil aggregation and water-holding capacity, whereas total EPS from anaerobic sludge have significant potential for ammonia removal under salt stress. Fourier transform infrared spectroscopy (FTIR) revealed that both EPS may contain proteins, carbohydrates, humic compounds, uronic acid, and DNA and determined the presence of O–H, N–H, C–N, CO, and C–H functional groups. These results demonstrate that the overall enzyme activity may be inactivated at 30 g L-1 of salinity. An annotation found in Kyoto Encyclopedia of Genes and Genomes (KEGG)- KEGG Automatic Annotation Server (KAAS) revealed that the top two metabolic activities in the EPS generated from the anaerobic sludge were methane and nitrogen metabolism. Therefore, we focused on the nitrogen metabolism reference map 00910. EPS from the anaerobically digested sludge exhibited nitrate reductase, nitrite reductase, and dehydrogenase activities. Assimilatory nitrate reduction, denitrification, nitrification, and anammox removed ammonia biochemically. The influence of microbial extracellular metabolites on water-holding capacity and soil aggregation was also investigated. The KAAS-KEGG annotation server was used to identify the main enzymes in the activated sludge-derived alginate-like extracellular EPS (ALE-EPS) samples. These include hydrolases, oxidoreductases, lyases, ligases, and transporters, which contribute to soil fertility and stability. This study improves our understanding of the overall microbial community structure and the associated biochemical processes, which are related to distinct functional genes or enzymes involved in nitrogen removal and soil aggregation. In contrast to conventional methods, microbial association with proteomics can be used to investigate ecological relationships, establishments, key player species, and microbial responses to environmental changes. Linking the metagenome to off-gel proteomics and bioinformatics solves the problem of analyzing metabolic pathways in complex environmental samples in a cost-effective manner.

One-pot production of 5-methylfurfural (5-MF) and enhanced dewaterability of waste activated sludge by hydrothermal treatment with natural deep eutectic solvents (NADES): Experimental and theoretical studies

Natural deep eutectic solvents (NADES) coupled with hydrothermal treatment was innovatively proposed to process waste activated sludge for sustainable production of 5-methylfurfural (5-MF) and enhanced dewaterability. The yield of 5-MF up to 11.54 kg/t volatile solids (VS) with a NADES dosage of 2.0 mL, hydrothermal temperature of 180 °C and holding time of 30 min. The decreased capillary suction time (CST) and bound water content indicated that NADES with hydrothermal treatment substantially improved the dewaterability of sludge. The variations of sludge physicochemical properties and extracellular polymeric substances (EPS) compositions revealed that NADES disrupted the structure of sludge biopolymers. Density functional theory (DFT) calculations indicated that NADES weakened the hydrogen bond strength between EPS components, and thus promoted the solubilization of proteins and polysaccharides. Besides, transition state analysis showed that NADES also lowered the energy barrier of polypeptide hydrolysis and carbohydrates dehydration reactions for the production of 5-MF.

Spectroscopic fingerprints profiling the polysaccharide/protein/humic architecture of stratified extracellular polymeric substances (EPS) in activated sludge

Extracellular polymeric substances (EPS), with a stratified structure including tightly-bound EPS (TB-EPS), loosely-bound EPS (LB-EPS), and soluble EPS (S-EPS) surrounding the microbial cells, are known to vitally affect the physicochemical and biological functions of activated sludge in wastewater treatment. Polysaccharides (PS), proteins (PN), and humic acids (HA) are key components of EPS but their roles in constructing the multi-layer architecture are still unclear. This study explored the EPS characteristics in relation to the components using spectroscopic fingerprinting techniques. Ultraviolet-visible (UV-vis) spectra demonstrated stark difference between TB-EPS and other EPS. Fluorescence excitation-emission matrix (FEEM) and apparent quantum yield revealed further detailed differences. Fluorescence quotient analysis highlighted the dominance of TB-EPS, LB-EPS, and S-EPS in the excitation/emission wavelength (Ex/Em) region of Em = 350–400 nm, Em > 400 nm, and low-Stokes shift band (Em − Ex < 25 nm), respectively. Wavelength-wise prediction of the FEEM intensity was achieved through multiple linear regression against the chemical composition and variance partitioning analysis witnessed binary interactions of PS×HA and PS×PN in S-EPS, PN×HA and PS×PN in LB-EPS, and ternary interaction of PS×PN×HA in TB-EPS as well as the wavelength-specific fluorescence responses of these interactions. Further, X-ray photoelectron spectroscopy, infrared spectra, and circular dichroism spectra corroborated the differences in primary, secondary, and tertiary structures across the EPS layers. Ultrahigh-performance liquid chromatography-mass spectrometry detected molecular fragments confirming the multi-component hybridization among PS, PN, and HA. This study demonstrates a spectroscopic approach to sensitively fingerprint the fine structure of EPS, which has the potential for rapid monitoring of EPS and related sludge properties in wastewater treatment systems.

Enhancement of biogranules development using magnetized powder activated carbon.

Biogranulation has emerged as a viable alternative biological wastewater treatment approach because of its strong biodegradability potential, toxicity tolerance, and biomass retention features. However, this process requires a long duration for biogranules formation to occur. In this study, magnetic powder activated carbon (MPAC) was used as support material in a sequencing batch reactor to enhance biogranules development for wastewater treatment. Two parallel SBRs (designated R1 and R2) were used, with R1 serving as a control without the presence of MPAC whileR2 wasoperated with MPAC. The biodegradability capacity and biomass properties of MPAC biogranules were compared with a control system. The measured diameter of biogranules for R1 and R2 after 8weeks of maturation were 2.2mm and 3.4mm, respectively. The integrity coefficient of the biogranules in R2 was higher (8.3%) than that of R1 (13.4%), indicating that the addition of MPAC improved the structure of the biogranules in R2. The components of extracellular polymeric substances were also higher in R2 than in R1. Scanning electronic microscopy was able to examine the morphological structures of the biogranules which showed there were irregular formations compacted together. However, there were more cavities situated in R1 biogranules (without MPAC) when compared to R2 biogranules (with MPAC). Dye removal reached 65% and 83% in R1 and R2 in the post-development stage. This study demonstrates that the addition of MPAC could shorten and improve biogranules formation. MPAC acted as the support media for microbial growth during the biogranulation developmental process.

A comparative study of the accumulation and detoxification of copper and zinc in Chlamydomonas reinhardtii: The role of extracellular polymeric substances

Extracellular polymeric substances (EPS) form an interface between microalgae and the surrounding water environment. Copper (Cu) and zinc (Zn) are essential micronutrients but may negatively affect microbial growth when their concentrations reach toxic thresholds. However, how EPS affect the accumulation and resistance of Cu and Zn in microalgae remains largely unknown. Here, we investigated EPS production upon Cu/Zn exposure and compared the tolerance strategies to the two metals by Chlamydomonas reinhardtii with and without EPS. Microalgal EPS synthesis was induced by Cu/Zn treatments, and the functional groups of polysaccharides and proteins were involved in complexation with metal ions. The extraction of EPS aggravated the toxicity and reduced the removal of metals from solution, but the effect was more pronounced for Cu than for Zn. Copper bound on the cell surface accounted for 54.6 ± 2.0 % of the Cu accumulated by C. reinhardtii, whose EPS components strongly correlated with Cu adsorption. In contrast, 74.3 ± 3.0 % of accumulated Zn was absorbed in cells, and glutathione synthesis was significantly induced. Redundancy and linear correlation analyses showed that the polysaccharide, protein and DNA contents in EPS were significantly correlated with Cu accumulation, absorption and adsorption but not with Zn. Data fitted to a Michaelis–Menten model further showed that the EPS-intact cells had higher binding capacity for Cu2+ but not for Zn2+. These differential impacts of EPS on Cu/Zn sorption and detoxification contribute to a more comprehensive understanding of the roles of microalgal EPS in the biogeochemical cycle of metals.

An RpoN-dependent PEP-CTERM gene is involved in floc formation of an Aquincola tertiaricarbonis strain

BackgroundThe floc is a characteristic of microbial aggregate growth, displaying cloudy suspensions in water. Floc formation has been demonstrated in a series of bacteria and the floc-forming bacteria play a crucial role in activated sludge (AS) process widely used for municipal sewage and industrial wastewater treatment over a century. It has been demonstrated that some exopolysaccharide biosynthesis genes and the sigma factor (sigma54 or rpoN) were required for floc forming in some bacteria. However, the mechanism underlying the floc formation stills need to be elucidated.ResultsIn this study, we demonstrate that a TPR (Tetratricopeptide repeats) protein-encoding gene prsT is required for floc formation of Aquincola tertiaricarbonis RN12 and an upstream PEP-CTERM gene (designated pepA), regulated by RpoN1, is involved in its floc formation but not swarming motility and biofilm formation. Overexpression of PepA could rescue the floc-forming phenotype of the rpoN1 mutant by decreasing the released soluble exopolysaccharides and increasing the bound polymers.ConclusionOur results indicate that the wide-spread PEP-CTERM proteins play an important role in the self-flocculation of bacterial cells and may be a component of extracellular polymeric substances required for floc-formation.

Molecular Dynamics Simulation of the Aggregation of Extracellular Polymeric Substance

Extracellular polymeric substance (EPS) play an important role in the transport and transformation of pollutants. At present, some scholars have conducted basic experimental characterization of EPS, but the molecular lev-el interactions among the various components of EPS still need to be fully characterized. Here, we use molecular dynamics (MD) simulation to explore the structural properties of EPS systems in atomic detail. The process and mechanism of the aggregation of different EPS components (polysaccharide, lipid, nucleic acid and protein) were revealed by the simulation results. EPS aggregation consist of a hydrophobic core and an amphiphilic exterior. Lipid tail, as a hydrophobic core, promoted the aggregation of EPS. But strong hy-drophilic nucleic acid and protein components inhibited the aggregation of EPS in water and were located outside the aggregation body. The details of the structure of EPS aggregation are revealed here, which provides a micro molecular mechanism for understanding the interaction between EPS.