ABSTRACT
As a significant structure in activated sludge, extracellular polymeric substances (EPS) hold considerable value regarding resource recovery and applications. The present study aimed to elucidate the relationship between the microbial community and the composition and properties of EPS. A biological nutrient removal (BNR) reactor was set up in the laboratory and controlled under different solid retention times (SRT), altering microbial species within the system. Then EPS was extracted from activated and analyzed by chemical and spectroscopic methods. High-throughput sequencing and metagenomic approaches were employed to investigate bacterial community and metabolic pathways. The results showed that lower SRT with a higher abundance of the family-level Proteobacteria (27.7%-53.5%) favored EPS synthesis, while another dominant group Bacteroidetes (20.0%-32.6%) may not significantly affect EPS synthesis. Furthermore, the abundance of alginates-producing bacteria including Pseudomonas spp. and Azotobacter vinelandii was only 2.53%-6.76% and 1.98%-6.34%, respectively. The alginate synthesis pathway genes Alg8 and Alg44 were also present at very low levels (0.05‱-0.11‱, 0.01‱-0.02‱, respectively). Another important gene related to alginates operons, AlgK, was absent across all the SRT-operated reactors. These findings suggest an impossible and incomplete alginate synthesis pathway within sludge. In light of these results, it can be concluded that EPS does not necessarily contain alginate components.
ABSTRACT
Biological wastewater treatment relies on microorganisms that grow as flocs, biofilms, or granules for efficient separation of biomass from cleaned water. This biofilm structure emerges from the interactions between microbes that produce, and are embedded in, extracellular polymeric substances (EPS). The true composition and structure of the EPS responsible for dense biofilm formation are still obscure. We conducted a bottom-up approach utilizing advanced glycomic techniques to explore the glycan diversity in the EPS from a highly enriched "Candidatus Accumulibacter" granular sludge. Rare novel sugar monomers such as N-Acetylquinovosamine (QuiNAc) and 2-O-Methylrhamnose (2-OMe-Rha) were identified to be present in the EPS of both enrichments. Further, a high diversity in the glycoprotein structures of said EPS was identified by means of lectin based microarrays. We explored the genetic potential of "Ca. Accumulibacter" high quality metagenome assembled genomes (MAGs) to showcase the shortcoming of top-down bioinformatics based approaches at predicting EPS composition and structure, especially when dealing with glycans and glycoconjugates. This work suggests that more bottom-up research is necessary to understand the composition and complex structure of EPS in biofilms since genome based inference cannot directly predict glycan structures and glycoconjugate diversity.
ABSTRACT
Biodegradable flocculants are rarely used in waste activated sludge (WAS) fermentation. This study introduces an alginate-based biodegradable flocculant (ABF) to enhance both the dewatering and degradation of WAS during its fermentation. Alginate was identified in structural extracellular polymeric substances (St-EPS) of WAS, with alginate-producing bacteria comprising â¼4.2% of the total bacterial population in WAS. Owing to its larger floc size, higher contact angle, and lower free energy resulting from the Lewis acid-base interaction, the addition of the prepared ABF with a network structure significantly improved the dewaterability of WAS and reduced capillary suction time (CST) by 72%. The utilization of ABF by an enriched alginate-degrading consortium (ADC) resulted in a 35.5% increase in the WAS methane yield owing to its higher hydrolytic activity on both ABF and St-EPS. Additionally, after a 30 day fermentation, CST decreased by 62% owing to the enhanced degradation of St-EPS (74.4%) and lower viscosity in the WAS + ABF + ADC group. The genus Bacteroides, comprising 12% of ADC, used alginate lyase (EC 4.2.2.3) and pectate lyase (EC 4.2.2.2 and EC 4.2.2.9) to degrade alginate and polygalacturonate in St-EPS, respectively. Therefore, this study introduces a new flocculant and elucidates its dual roles in enhancing both the dewaterability and degradability of WAS. These advancements improve WAS fermentation, resulting in higher methane production and lower CSTs.
Subject(s)
Alginates , Fermentation , Flocculation , Sewage , Anaerobiosis , Waste Disposal, Fluid , Biodegradation, EnvironmentalABSTRACT
Wastewater treatment technologies opened the door for recovery of extracellular polymeric substances (EPS), presenting novel opportunities for use across diverse industrial sectors. Earlier studies showed that a significant amount of phosphorus (P) is recovered within extracted EPS. P recovered within the extracted EPS is an intrinsic part of the recovered material that potentially influences its properties. Understanding the P speciation in extracted EPS lays the foundation for leveraging the incorporated P in EPS to manipulate its properties and industrial applications. This study evaluated P speciation in EPS extracted from aerobic granular sludge (AGS). A fractionation lab protocol was established to consistently distinguish P species in extracted EPS liquid phase and polymer chains. 31P nuclear magnetic resonance (NMR) spectroscopy was used as a complementary technique to provide additional information on P speciation and track changes in P species during the EPS extraction process. Findings showed the dominance of organic phosphorus and orthophosphates within EPS, besides other minor fractions. On average, 25% orthophosphates in the polymer liquid phase, 52% organic phosphorus (equal ratio of mono and diesters) covalently bound to the polymer chains, 16% non-apatite inorganic phosphorus (NAIP) precipitates mainly FeP and AlP, and 7% pyrophosphates (6% in the liquid phase and 1% attached to the polymer chains) were identified. Polyphosphates were detected in initial AGS but hydrolyzed to orthophosphates, pyrophosphates, and possibly organic P (forming new esters) during the EPS extraction process. The knowledge created in this study is a step towards the goal of EPS engineering, manipulating P chemistry along the extraction process and enriching certain P species in EPS based on target properties and industrial applications.
Subject(s)
Extracellular Polymeric Substance Matrix , Phosphorus , Sewage , Sewage/chemistry , Extracellular Polymeric Substance Matrix/chemistry , Magnetic Resonance Spectroscopy , AerobiosisABSTRACT
Rapid sand filters are established and widely applied technologies for groundwater treatment. In these filters, main groundwater contaminants such as iron, manganese, and ammonium are oxidized and removed. Conventionally, intensive aeration is employed to provide oxygen for these redox reactions. While effective, intensive aeration promotes flocculent iron removal, which results in iron oxide flocs that rapidly clog the filter. In this study, we operated two parallel full-scale sand filters at different aeration intensities to resolve the relative contribution of homogeneous, heterogeneous and biological iron removal pathways, and identify their operational controls. Our results show that mild aeration in the LOW filter (5 mg/L O2, pH 6.9) promoted biological iron removal and enabled iron oxidation at twice the rate compared to the intensively aerated HIGH filter (>10 mg/L O2, pH 7.4). Microscopy images showed distinctive twisted stalk-like iron solids, the biosignatures of Gallionella ferruginea, both in the LOW filter sand coatings as well as in its backwash solids. In accordance, 10 times higher DNA copy numbers of G. ferruginea were found in the LOW filter effluent. Clogging by biogenic iron solids was slower than by chemical iron flocs, resulting in lower backwash frequencies and yielding four times more water per run. Ultimately, our results reveal that biological iron oxidation can be actively controlled and favoured over competing physico-chemical routes. The production of more compact and practically valuable iron oxide solids is of outmost interest. We conclude that, although counterintuitive, slowing down iron oxidation in the water before filtration enables rapid iron removal in the biofilter.
Subject(s)
Filtration , Groundwater , Iron , Water Purification , Groundwater/chemistry , Iron/chemistry , Water Purification/methods , Water Pollutants, Chemical , Gallionellaceae , Oxidation-ReductionABSTRACT
Nitrous oxide (N2O) is a potent greenhouse gas of primarily microbial origin. Oxic and anoxic emissions are commonly ascribed to autotrophic nitrification and heterotrophic denitrification, respectively. Beyond this established dichotomy, we quantitatively show that heterotrophic denitrification can significantly contribute to aerobic nitrogen turnover and N2O emissions in complex microbiomes exposed to frequent oxic/anoxic transitions. Two planktonic, nitrification-inhibited enrichment cultures were established under continuous organic carbon and nitrate feeding, and cyclic oxygen availability. Over a third of the influent organic substrate was respired with nitrate as electron acceptor at high oxygen concentrations (>6.5 mg/L). N2O accounted for up to one-quarter of the nitrate reduced under oxic conditions. The enriched microorganisms maintained a constitutive abundance of denitrifying enzymes due to the oxic/anoxic frequencies exceeding their protein turnover-a common scenario in natural and engineered ecosystems. The aerobic denitrification rates are ascribed primarily to the residual activity of anaerobically synthesised enzymes. From an ecological perspective, the selection of organisms capable of sustaining significant denitrifying activity during aeration shows their competitive advantage over other heterotrophs under varying oxygen availabilities. Ultimately, we propose that the contribution of heterotrophic denitrification to aerobic nitrogen turnover and N2O emissions is currently underestimated in dynamic environments.
Subject(s)
Bacteria , Denitrification , Microbiota , Nitrous Oxide , Nitrous Oxide/metabolism , Aerobiosis , Bacteria/metabolism , Bacteria/genetics , Bacteria/classification , Heterotrophic Processes , Nitrates/metabolism , Oxygen/metabolism , Nitrogen/metabolism , NitrificationABSTRACT
Phosphorous not only needs to be removed to prevent eutrophication of wastewater effluent receiving surface water bodies, but it also has to be recovered as a scarce finite reserve. Phosphorus chemical precipitation as NH4MgPO4·6H2O, Ca3(PO4)2, or Fe3(PO4)2 ·8H2O is the most common method of phosphorus recovery from phosphorus-rich streams. These minerals ideally form under neutral to alkaline pH conditions, making acidic streams problematic for their formation due to the need for pH adjustments. This study proposes FePO4 .2H2O (strengite-like compounds) recovery from acidic streams due to its simplicity and high efficiency, while also avoiding the need for pH-adjusting chemicals. The effect of initial pH, temperature, Fe (III) dosing rates, and Fe (II) dosage under different oxidation conditions (pO2 = 0.2, 1, 1.5 bar, different H2O2 dosing rates) on phosphorus recovery percentage and product settleability were evaluated in this study. The precipitates formed were analyzed using optical microscopy, SEM, XRD, SQUID, Raman, and ICP. Experiments showed that Fe (III) dosing achieved phosphorus recovery of over 95 % at an initial pH of 3 or higher, and the product exhibited poor settleability in all initial pH (1.5-5), and temperature (20-80 °C) tests. On the other hand, Fe (II) dosage instead of Fe (III) resulted in good product settleability but varying phosphorus recovery percentages depending on the oxidation conditions. The novelty of the study lies in revealing that the Fe (II) oxidation rate serves as a crucial process-design parameter, significantly enhancing product settleability without the requirement of carrier materials or crystallizers. The study proposes a novel strategy with controlled Fe2+-H2O2 dosing, identifying an Fe (II) oxidation rate of 4.7 × 10-4 mol/l/min as the optimal rate for achieving over 95 % total phosphorus recovery, along with excellent settleability with a volumetric index equal to only 8 ml/gP.
Subject(s)
Phosphates , Phosphorus , Phosphates/chemistry , Hydrogen-Ion Concentration , Phosphorus/chemistry , Wastewater/chemistry , Waste Disposal, Fluid/methods , Hydrogen Peroxide/chemistry , Ferric Compounds/chemistry , Chemical PrecipitationABSTRACT
Rapid sand filters (RSF) are an established and widely applied technology for the removal of dissolved iron (Fe2+) and ammonium (NH4+) among other contaminants in groundwater treatment. Most often, biological NH4+oxidation is spatially delayed and starts only upon complete Fe2+ depletion. However, the mechanism(s) responsible for the inhibition of NH4+oxidation by Fe2+ or its oxidation (by)products remains elusive, hindering further process control and optimization. We used batch assays, lab-scale columns, and full-scale filter characterizations to resolve the individual impact of the main Fe2+ oxidizing mechanisms and the resulting products on biological NH4+ oxidation. modeling of the obtained datasets allowed to quantitatively assess the hydraulic implications of Fe2+ oxidation. Dissolved Fe2+ and the reactive oxygen species formed as byproducts during Fe2+ oxidation had no direct effect on ammonia oxidation. The Fe3+ oxides on the sand grain coating, commonly assumed to be the main cause for inhibited ammonia oxidation, seemed instead to enhance it. modeling allowed to exclude mass transfer limitations induced by accumulation of iron flocs and consequent filter clogging as the cause for delayed ammonia oxidation. We unequivocally identify the inhibition of NH4+oxidizing organisms by the Fe3+ flocs generated during Fe2+ oxidation as the main cause for the commonly observed spatial delay in ammonia oxidation. The addition of Fe3+ flocs inhibited NH4+oxidation both in batch and column tests, and the removal of Fe3+ flocs by backwashing completely re-established the NH4+removal capacity, suggesting that the inhibition is reversible. In conclusion, our findings not only identify the iron form that causes the inhibition, albeit the biological mechanism remains to be identified, but also highlight the ecological importance of iron cycling in nitrifying environments.
Subject(s)
Filtration , Groundwater , Iron , Nitrification , Groundwater/chemistry , Iron/chemistry , Ammonium Compounds , Oxidation-Reduction , Ammonia , Water Purification/methodsABSTRACT
Industrial wastewater often has high levels of salt, either due to seawater or e.g. sodium chloride (NaCl) usage in the processing. Previous work indicated that aerobic granular sludge (AGS) is differently affected by seawater or saline water at similar osmotic strength. Here we investigate in more detail the impact of NaCl concentrations and seawater on the granulation and conversion processes for AGS wastewater treatment. Glycerol was used as the carbon source since it is regularly present in industrial wastewaters, and to allow the evaluation of microbial interactions that better reflect real conditions. Long-term experiments were performed to evaluate and compare the effect of salinity on granulation, anaerobic conversions, phosphate removal, and the microbial community. Smooth and stable granules as well as enhanced biological phosphorus removal (EBPR) were achieved up to 20 g/L NaCl or when using seawater. However, at NaCl levels comparable to seawater strength (30 g/L) incomplete anaerobic glycerol uptake and aerobic phosphate uptake were observed, the effluent turbidity increased, and filamentous granules began to appear. The latter is likely due to the direct aerobic growth on the leftover substrate after the anaerobic feeding period. In all reactor conditions, except the reactor with 30 g/L NaCl, Ca. Accumulibacter was the dominant microorganism. In the reactor with 30 g/L NaCl, the relative abundance of Ca. Accumulibacter decreased to ≤1 % and an increase in the genus Zoogloea was observed. Throughout all reactor conditions, Tessaracoccus and Micropruina, both actinobacteria, were present which were likely responsible for the anaerobic conversion of glycerol into volatile fatty acids. None of the glycerol metabolizing proteins were detected in Ca. Accumulibacter which supports previous findings that glycerol can not be directly utilized by Ca. Accumulibacter. The proteome profile of the dominant taxa was analysed and the results are further discussed. The exposure of salt-adapted biomass to hypo-osmotic conditions led to significant trehalose and PO43--P release which can be related to the osmoregulation of the cells. Overall, this study provides insights into the effect of salt on the operation and stability of the EBPR and AGS processes. The findings suggest that maintaining a balanced cation ratio is likely to be more important for the operational stability of EBPR and AGS systems than absolute salt concentrations.
Subject(s)
Glycerol , Phosphorus , Salinity , Sewage , Sewage/microbiology , Phosphorus/metabolism , Glycerol/metabolism , Aerobiosis , Bioreactors , Waste Disposal, FluidABSTRACT
Aerobic granular sludge (AGS) and conventional activated sludge (CAS) are two different biological wastewater treatment processes. AGS consists of self-immobilised microorganisms that are transformed into spherical biofilms, whereas CAS has floccular sludge of lower density. In this study, we investigated the treatment performance and microbiome dynamics of two full-scale AGS reactors and a parallel CAS system at a municipal WWTP in Sweden. Both systems produced low effluent concentrations, with some fluctuations in phosphate and nitrate mainly due to variations in organic substrate availability. The microbial diversity was slightly higher in the AGS, with different dynamics in the microbiome over time. Seasonal periodicity was observed in both sludge types, with a larger shift in the CAS microbiome compared to the AGS. Groups important for reactor function, such as ammonia-oxidising bacteria (AOB), nitrite-oxidising bacteria (NOB), polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs), followed similar trends in both systems, with higher relative abundances of PAOs and GAOs in the AGS. However, microbial composition and dynamics differed between the two systems at the genus level. For instance, among PAOs, Tetrasphaera was more prevalent in the AGS, while Dechloromonas was more common in the CAS. Among NOB, Ca. Nitrotoga had a higher relative abundance in the AGS, while Nitrospira was the main nitrifier in the CAS. Furthermore, network analysis revealed the clustering of the various genera within the guilds to modules with different temporal patterns, suggesting functional redundancy in both AGS and CAS. KEY POINTS: ⢠Microbial community succession in parallel full-scale aerobic granular sludge (AGS) and conventional activated sludge (CAS) processes. ⢠Higher periodicity in microbial community structure in CAS compared to in AGS. ⢠Similar functional groups between AGS and CAS but different composition and dynamics at genus level.
Subject(s)
Bacteria , Bioreactors , Microbiota , Sewage , Sewage/microbiology , Bacteria/classification , Bacteria/metabolism , Bacteria/genetics , Bacteria/isolation & purification , Bioreactors/microbiology , Aerobiosis , Sweden , Glycogen/metabolism , Ammonia/metabolism , Nitrites/metabolism , Nitrates/metabolism , Phosphates/metabolism , Water Purification/methodsABSTRACT
Reverse osmosis (RO) systems offer a viable solution for treating brackish water (BW), a common but underutilized water resource. However, the energy-intensive nature of brackish water reverse osmosis (BWRO) systems poses affordability challenges to water supply, necessitating a focus on minimizing their energy consumption to support SDG6's goal of providing safe and affordable drinking water for all. This study addresses the critical need to minimize the specific energy consumption (SEC) of a typical BWRO system, defined as the energy consumed per unit of water recovered, mathematically and experimentally. Empirical models were developed proving there is a global minimum SEC while adjusting the operating conditions. Furthermore, we identified the key operating factors influencing SEC and their priority levels, along with their interactive effects. Notably, no prior study has discussed the significance and interaction of these operating factors (e.g., feed water salinity, temperature, pressure, flowrate and membrane permeability) on SEC of a BWRO system. Employing a full factorial experimental design with mixed levels of operating parameters, the study developed regression models that elucidate the mechanistic interaction between these parameters and system performance. Moreover, the models were validated experimentally, with a new dataset demonstrating their accuracy and reliability. ANOVA statistical analysis identified feed salinity, pressure, flow rate, feed flow rate×pressure, salinity×pressure, and temperature as influential operating parameters in reducing SEC, in descending order of importance. Operating within the determined optimum range resulted in a 36 % decrease in SEC and a more than fourfold increase in water recovery. The study's systematic approach and findings can be extrapolated to optimize the performance of other desalination technologies and diverse feed water types, contributing significantly to global water sustainability efforts.
ABSTRACT
Nitrate leaching from agricultural soils is increasingly found in groundwater, a primary source of drinking water worldwide. This nitrate influx can potentially stimulate the biological oxidation of iron in anoxic groundwater reservoirs. Nitrate-dependent iron-oxidizing (NDFO) bacteria have been extensively studied in laboratory settings, yet their ecophysiology in natural environments remains largely unknown. To this end, we established a pilot-scale filter on nitrate-rich groundwater to elucidate the structure and metabolism of nitrate-reducing iron-oxidizing microbiomes under oligotrophic conditions mimicking natural groundwaters. The enriched community stoichiometrically removed iron and nitrate consistently with the NDFO metabolism. Genome-resolved metagenomics revealed the underlying metabolic network between the dominant iron-dependent denitrifying autotrophs and the less abundant organoheterotrophs. The most abundant genome belonged to a new Candidate order, named Siderophiliales. This new species, "Candidatus Siderophilus nitratireducens," carries genes central genes to iron oxidation (cytochrome c cyc2), carbon fixation (rbc), and for the sole periplasmic nitrate reductase (nap). Using thermodynamics, we demonstrate that iron oxidation coupled to nap based dissimilatory reduction of nitrate to nitrite is energetically favorable under realistic Fe3+/Fe2+ and NO3-/NO2- concentration ratios. Ultimately, by bridging the gap between laboratory investigations and nitrate real-world conditions, this study provides insights into the intricate interplay between nitrate and iron in groundwater ecosystems, and expands our understanding of NDFOs taxonomic diversity and ecological role.
ABSTRACT
Glycans are crucial for the structure and function of anaerobic granular sludge in wastewater treatment. Yet, there is limited knowledge regarding the microorganisms and biosynthesis pathways responsible for glycan production. In this study, we analysed samples from anaerobic granular sludges treating papermill and brewery wastewater, examining glycans composition and using metagenome-assembled genomes (MAGs) to explore potential biochemical pathways associated with their production. Uronic acids were the predominant constituents of the glycans in extracellular polymeric substances (EPS) produced by the anaerobic granular sludges, comprising up to 60 % of the total polysaccharide content. MAGs affiliated with Anaerolineacae, Methanobacteriaceae and Methanosaetaceae represented the majority of the microbial community (30-50 % of total reads per MAG). Based on the analysis of MAGs, it appears that Anaerolinea sp. and members of the Methanobacteria class are involved in the production of exopolysaccharides within the analysed granular sludges. These findings shed light on the functional roles of microorganisms in glycan production in industrial anaerobic wastewater treatment systems.
Subject(s)
Metagenome , Sewage , Sewage/chemistry , Anaerobiosis , Wastewater , Polysaccharides , Bioreactors/microbiology , Waste Disposal, FluidABSTRACT
The accumulation of phosphorus in activated sludge in wastewater treatment plants (WWTPs) provides potential for phosphorus recovery from sewage. This study delves into the potential for releasing phosphorus from waste activated sludge through two distinct treatment methods-thermal hydrolysis and pH adjustment. The investigation was conducted with activated sludge sourced from four WWTPs, each employing distinct phosphorus removal strategies. The findings underscore the notably superior efficacy of pH adjustment in solubilizing sludge phosphorus compared to the prevailing practice of thermal hydrolysis, widely adopted to enhance sludge digestion. The reversibility of phosphorus release within pH fluctuations spanning 2 to 12 implies that the release of sludge phosphorus can be attributed to the dissolution of phosphate precipitates. Alkaline sludge treatment induced the concurrent liberation of COD, nitrogen, and phosphorus through alkaline hydrolysis of sludge biomass and the dissolution of iron or aluminium phosphates, offering potential gains in resource recovery and energy efficiency.
Subject(s)
Wastewater , Water Purification , Sewage , Phosphorus , Nitrogen , Carbon , Waste Disposal, FluidABSTRACT
Currently, the most cost-effective and efficient method for phosphorus (P) removal from wastewater is enhanced biological P removal (EPBR) via polyphosphate-accumulating organisms (PAOs). This study integrates a literature review with genomic analysis to uncover the phylogenetic and metabolic diversity of the relevant PAOs for wastewater treatment. The findings highlight significant differences in the metabolic capabilities of PAOs relevant to wastewater treatment. Notably, Candidatus Dechloromonas and Candidatus Accumulibacter can synthesize polyhydroxyalkanoates, possess specific enzymes for ATP production from polyphosphate, and have electrochemical transporters for acetate and C4-dicarboxylates. In contrast, Tetrasphaera, Candidatus Phosphoribacter, Knoellia, and Phycicoccus possess PolyP-glucokinase and electrochemical transporters for sugars/amino acids. Additionally, this review explores various detection methods for polyphosphate and PAOs in activated sludge wastewater treatment plants. Notably, FISH-Raman spectroscopy emerges as one of the most advanced detection techniques. Overall, this review provides critical insights into PAO research, underscoring the need for enhanced strategies in biological phosphorus removal.
ABSTRACT
This study investigates the development of polyvinyl alcohol (PVA) gel matrices for biomass immobilization in wastewater treatment. The PVA hydrogels were prepared through a freezing-thawing (F-T) cross-linking process and reinforced with high surface area nanoparticles to improve their mechanical stability and porosity. The PVA/nanocomposite hydrogels were prepared using two different nanoparticle materials: iron oxide (Fe3O2) and titanium oxide (TiO2). The effects of the metal oxide nanoparticle type and content on the pore structure, hydrogel bonding, and mechanical and viscoelastic properties of the cross-linked hydrogel composites were investigated. The most durable PVA/nanoparticles matrix was then tested in the bioreactor for the biological treatment of wastewater. Morphological analysis showed that the reinforcement of PVA gel with Fe2O3 and TiO2 nanoparticles resulted in a compact nanocomposite hydrogel with regular pore distribution. The FTIR analysis highlighted the formation of bonds between nanoparticles and hydrogel, which caused more interaction within the polymeric matrix. Furthermore, the mechanical strength and Young's modulus of the hydrogel composites were found to depend on the type and content of the nanoparticles. The most remarkable improvement in the mechanical strength of the PVA/nanoparticles composites was obtained by incorporating 0.1 wt% TiO2 and 1.0 wt% Fe2O3 nanoparticles. However, TiO2 showed more influence on the mechanical strength, with more than 900% improvement in Young's modulus for TiO2-reinforced PVA hydrogel. Furthermore, incorporating TiO2 nanoparticles enhanced hydrogel stability but did not affect the biodegradation of organic pollutants in wastewater. These results suggest that the PVA-TiO2 hydrogel has the potential to be used as an effective carrier for biomass immobilization and wastewater treatment.
ABSTRACT
Bacteria can synthesize a diverse array of glycans, being found attached to proteins and lipids or as loosely associated polysaccharides to the cells. The major challenge in glycan analysis in environmental samples lies in developing high-throughput and comprehensive characterization methodologies to elucidate the structure and monitor the change of the glycan profile, especially in protein glycosylation. To this end, in the current research, the dynamic change of the glycan profile of a few extracellular polymeric substance (EPS) samples was investigated by high-throughput lectin microarray and mass spectrometry, as well as sialylation and sulfation analysis. Those EPS were extracted from aerobic granular sludge collected at different stages during its adaptation to the seawater condition. It was found that there were glycoproteins in all of the EPS samples. In response to the exposure to seawater, the amount of glycoproteins and their glycan diversity displayed an increase during adaptation, followed by a decrease once the granules reached a stable state of adaptation. Information generated sheds light on the approaches to identify and monitor the diversity and dynamic alteration of the glycan profile of the EPS in response to environmental stimuli.
ABSTRACT
Anionic polymers, such as heparin, have been widely applied in the chemical and medical fields, particularly for binding proteins (e.g., fibroblast growth factor 2 (FGF-2) and histones). However, the current animal-based production of heparin brings great risks, including resource shortages and product contamination. Recently, anionic compounds, nonulosonic acids (NulOs), and sulfated glycoconjugates were discovered in the extracellular polymeric substances (EPS) of aerobic granular sludge (AGS). Given the prevalence of anionic polymers, in marine biofilms, it was hypothesized that the EPS from AGS grown under seawater condition could serve as a raw material for producing the alternatives to heparin. This study aimed to isolate and enrich the anionic fractions of EPS and evaluate their potential application in the chemical and medical fields. The AGS was grown in a lab-scale reactor fed with acetate, under the seawater condition (35 g/L sea salt). The EPS was extracted with an alkaline solution at 80 °C and fractionated by size exclusion chromatography. Its protein binding capacity was evaluated by native gel electrophoresis. It was found that the two highest molecular weight fractions (438- > 14,320 kDa) were enriched with NulO and sulfate-containing glycoconjugates. The enriched fractions can strongly bind the two histones involved in sepsis and a model protein used for purification by heparin-column. These findings demonstrated possibilities for the application of the extracted EPS and open up a novel strategy for resource recovery. KEY POINTS: ⢠High MW EPS from seawater-adapted AGS are dominant with sulfated groups and NulOs ⢠Fifty-eight percent of the EPS is high MW of 68-14,320 kDa ⢠EPS and its fractions can bind histones and fibroblast growth factor 2.