Role Of Extracellular Polymeric Substances Research Articles

Deciphering the role of extracellular polymeric substances in the adsorption and biotransformation of organic micropollutants during anaerobic wastewater treatment

Extracellular polymeric substances (EPS) participate in the removal of organic micropollutants (OMPs), but the primary pathways of removal and detailed mechanisms remain elusive. We evaluated the effect of EPS on removal for 16 distinct chemical classes of OMPs during anaerobic digestion (AD). The results showed that hydrophobic OMPs (HBOMPs) could not be removed by EPS, while hydrophilic OMPs (HLOMPs) were amenable to removal via adsorption and biotransformation of EPS. The adsorption and biotransformation of HLOMPs by EPS accounted up to 19.4 ± 0.9 % and 6.0 ± 0.8 % of total removal, respectively. Further investigations into the adsorption and biotransformation mechanisms of HLOMPs by EPS were conducted utilizing spectral, molecular dynamics simulation, and electrochemical analysis. The results suggested that EPS provided abundant binding sites for the adsorption of HLOMPs. The binding of HLOMPs to tryptophan-like proteins in EPS formed nonfluorescent complexes. Hydrogen bonds, hydrophobic interactions and water bridges were key to the binding processes and helped stabilize the complexes. The biotransformation of HLOMPs by EPS may be attributed to the presence of extracellular redox active components (c-type cytochromes (c-Cyts), c-Cyts-bound flavins). This study enhanced the comprehension for the role of EPS on the OMPs removal in anaerobic wastewater treatment.

Harnessing the Potential of Extracellular Polymeric Substances in Enhancing ANAMMOX Processes: Mechanisms, Strategies, and Perspectives

Anaerobic ammonium oxidation (ANAMMOX) has emerged as a promising sustainable nitrogen removal technology that offers significant advantages over conventional nitrification–denitrification processes, such as reduced energy consumption, a 60% reduction in oxygen demand, and a 90% reduction in sludge production. However, the practical application of ANAMMOX is hindered by several challenges, including the slow growth of ANAMMOX bacteria, long start-up periods, and high sensitivity to environmental disturbances. Recent studies have highlighted the crucial role of extracellular polymeric substances (EPSs) in the formation, activity, and stability of ANAMMOX biofilms and granules. An EPS is a complex mixture of high-molecular-weight polymers secreted by microorganisms, mainly composed of polysaccharides, proteins, nucleic acids, and lipids. The diverse physicochemical properties and functional groups of EPSs enable them to serve as a structural scaffold, protective barrier, sorption site, electron shuttle, and nutrient source for ANAMMOX bacteria. This review aims to provide an overview of the latest research progress on harnessing the potential of EPSs to enhance the ANAMMOX process. The characteristics, compositions, and extraction methods of ANAMMOX-derived EPSs are summarized. The mechanisms of how EPSs facilitate the enrichment, immobilization, aggregation, and adaptation of ANAMMOX bacteria are elucidated. The strategies and effects of EPS supplementation on improving the performance and robustness of ANAMMOX reactors under various stresses are critically reviewed. The challenges and future perspectives of the EPS-mediated optimization of the ANAMMOX process are also discussed. This review sheds new light on exploiting EPSs as a renewable bioresource to develop more efficient and stable ANAMMOX applications for sustainable wastewater treatment.

Evaluating the feasibility of carbon fiber brush for improving biohydrogen production in acidogenic dark fermentation

Dark fermentation is a promising technology for biohydrogen (bioH2) production. Augmenting the dark fermentation process with conductive materials has drawn significant research interests improving the bioH2 yield. This study evaluated the feasibility of using carbon fiber brush (CB) as an additive to increase the efficiency of mesophilic dark fermentation using glucose as substrate and heat-shock pretreated anaerobic sludge as inoculum. Effect of electrode packing density on bioH2 yield, soluble metabolite product formation, and substrate degradation was investigated. Maximal bioH2 yield of 334.6 mL/g-glucose was obtained using medium-sized CB with 72.5% higher than the control (194 mL/g-glucose) and butyrate-type fermentation was the dominant metabolic pathway. Further, enhanced concentrations of proteins and polysaccharides on CB biofilm suggested the critical role of extracellular polymeric substances in microbial electron transfer. This was supported via electron transport activity in CB biofilm. Finally, microbial community analysis revealed a selective enrichment of the potentially electroactive Clostridium_sensu_stricto_1 on the CB (56.6%) compared to 40.9% abundance in control. Insights gained from this study demonstrated that CB aided dark fermentation can be an economical and environmentally benign strategy for improving bioH2 yield.

Roles of extracellular polymeric substances in arsenic accumulation and detoxification by cell wall intact and mutant strains of Chlamydomonas reinhardtii

Arsenic (As) pollution seriously threatens human and ecological health. Microalgal cell wall and extracellular polymeric substances (EPS) are known to interact with As, but their roles in the As resistance, accumulation and speciation in microalgae remain unclear. Here, we used two strains of Chlamydomonas reinhardtii, namely CC-125 (wild type) and CC-503 (cell wall-deficient mutant), to examine the algal growth, EPS synthesis, As adsorption, absorption and transformation under 10–1000 µg/L As(III) and As(V) treatments for 96 h. In both strains, the As absorption increased after the EPS removal, but the growth, As adsorption, and transformation of C. reinhardtii declined. The CC-125 strain was more tolerant to As stress and more efficient in EPS production, As accumulation, and redox transformation than CC-503, irrespective of EPS presence or absence. Three-dimension excitation-emission matrix (3D-EEM) and attenuated total reflectance infrared spectroscopy (ATR-IR) analyses showed that As was bound with functional groups in the EPS and cell wall, such as -COOH, -NH and -OH in proteins, polysaccharides and amino acids. Together, this study demonstrated that EPS and cell wall acted as barriers to lower the As uptake by C. reinhardtii. However, the cell wall mutant strain was more susceptible to As toxicity due to lower EPS induction and higher As absorption.

Insights into the degradation of diesel-contaminated seawater through natural microbiota and culturable microorganisms: The overlook role of extracellular polymeric substances (EPS)

The role of EPS produced during biodegradation diesel-contaminated seawater, as well as the effect of EPS on natural microbiota and culturable microorganisms, remains unknown. Here, natural microbiota (pre and post enrichment with diesel) from 5 islands’ seawater in Jiaozhou Bay were explored through metagenomic and culture-dependent approaches. EPS response degradation was mainly characterized by the following aspects: 1) The post-enrichment samples contained significantly fewer dominant bacterial genera than the pre-enrichment samples, and EPS or PS (polysaccharides) promoted degradation of C16-C24 hydrocarbon fractions; 2) EPS caused higher increases in aromatic compound degradation functional genes copies; 3) EPS enhanced the enzymatic activity of alkane hydroxylase and ethanol dehydrogenase; 4) Diesel degradation by mono and mixed cultures of culturable bacteria (B1 and B2) with extracted EPS showed a higher biomass, biosurfactant production, expression of PAH-RHDα GN and cytochrome P450 genes, degradability and emulsification capacity. The above innovations provided new perspectives for oil-spill bioremediation.

Role of extracellular polymeric substances in adsorptive binding of heavy metals on to Microsystis aeruginosa

Role of extracellular polymeric substances in adsorptive binding of heavy metals on to Microsystis aeruginosa

Role of extracellular polymeric substances in selenite and tellurite reduction by waste activated and anaerobic sludge

This study investigated the (simultaneous) reduction of selenite and tellurite by the Extracellular Polymeric Substances (EPS) of waste activated and anaerobic granular sludge. The EPS reduced the oxyanions to their...

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.

Removal of iron, manganese and ammonium from contaminated mine water by biochar immobilized Acinetobacter sp. AL-6 under acid condition, and the role of extracellular polymeric substances

In acidic mine wastewater, Mn and Fe are among the prime heavy metals and pose a great threat to the ecological environment and human health. Combining biological and physical methods to treat heavy metal contaminated water bodies is a promising remediation technique. In this study, grapefruit peel biochar immobilized Acinetobacter sp. AL-6 (co-system) was applied to treat the contaminated water with co-existing Mn2+, Fe2+ and NH4+-N. The results showed that almost 100% of Mn2+, Fe2+, Zn2+ and Cu2+ were removed in bioreactor, and was transferred to Mn2+, Fe3+, Zn2+ and Cu2+ precipitates on the surface of co-system without N2O accumulation. Especially, Mn2+ and Fe2+ were adsorbed in extracellular polymers (EPS) of strain AL-6. The present study mainly confirmed that the combined system of biochar and bacteria had great potential in the complex water removal of multiple heavy metals and NH4+-N, and also revealed the regulation of heavy metals by the EPS of strain.

Eco-corona formation diminishes the cytogenotoxicity of graphene oxide on Allium cepa: Role of soil extracted–extracellular polymeric substances in combating oxidative stress

Graphene oxide (GO) is widely acknowledged for its exceptional biological and industrial applications. However, its discharge into the environment negatively impacts the ecosystem. This study aimed to investigate the toxicity of GO in Allium cepa root tip cells and the role of extracellular polymeric substances (EPS) in modulating its toxic effects. To evaluate toxicity, various endpoints like cell viability using Evans blue dye, cytotoxicity (mitotic index), genotoxicity (chromosomal aberrations), and oxidative stress assessments (total ROS, superoxide, hydroxyl radical production, and lipid peroxidation) were considered. The results suggest that pristine GO caused a dose-dependent increase in various toxicity parameters, especially the genotoxic effects. Oxidative stress generation by GO is proposed to be the principal mode of action. The EPS-corona formed on GO could potentially counteract the toxic effects, substantially reducing the oxidative stress within the cells.

The influence of different nitrate concentrations on aerobic sludge granulation and the role of extracellular polymeric substances

This study investigated the influence of nitrate on aerobic granular sludge (AGS) granulation. The introduction of nitrate at 5, 15 and 20 mg L−1 promoted AGS granulation, and the promoting effect was positively correlated with nitrate concentrations. Meanwhile, exogenous nitrate significantly increased denitrification rate in the AGS system. However, granular disintegration appeared at a long-term addition of nitrate. An in-deep analysis showed that nitrate stimulated the secretion of extracellular polymeric substances (EPS), especially the content of proteins, which might be the main reason for the AGS granulation. However, the rapid and excessive increase in EPS might cause granular disintegration, as excessive EPS blocked the transmission of substrates, leading to the increase of dead cells in the granules. Besides, nitrate also altered the hydrophobicity of EPS and the content of α-helix, 3-turned helix and polymeric chain that favored aggregation, which also affected AGS granulation. From the microbial community level, nitrate induced the enrichment of denitrifying bacteria, including those that also functioned as EPS producers, such as Micropruina and Flavobacterium, resulting in the rapid increase of functional enzymes associated with amino acid synthesis, thereby promoting the secretion of proteins in EPS. Conversely, disintegration caused by mass transfer blockage might lead to the loss of EPS producing bacteria and subsequent decrease in EPS content, further accelerating granular disintegration.

Role of extracellular polymeric substances in resistance to allelochemical stress on Microcystis aeruginsosa and its mechanism

Role of extracellular polymeric substances in resistance to allelochemical stress on Microcystis aeruginsosa and its mechanism

Indole-3-acetic acid mediated removal of sludge toxicity by microalgae: Focus on the role of extracellular polymeric substances

The use of indole-3-acid (IAA) as an additive aided in achieving the objectives of reducing sludge extract toxicity, increasing Tetradesmus obliquus biomass yield, and enhancing extracellular polysaccharide production. Proteomics analysis can unveil the microalgae’s response mechanism to sludge toxicity stress. With 10-6 M IAA addition, microalgae biomass reached 3.426 ± 0.067 g/L. Sludge extract demonstrated 78.3 ± 3.2% total organic carbon removal and 72.2 ± 2.1% toxicity removal. Extracellular polysaccharides and proteins witnessed 2.08 and 1.76-fold increments, respectively. Proteomic analysis indicated that Tetradesmus obliquus directed carbon sources towards glycogen accumulation and amino acid synthesis, regulating pathways associated with carbon metabolism (glycolysis, TCA cycle, and amino acid metabolism) to adapt to the stressful environment. These findings lay the groundwork for future waste sludge treatment and offer novel insights into microalgae cultivation and extracellular polysaccharide enrichment in sludge.

Stable Aggregate Formation and Microbial Diversity Resilience in Soil Formation of Bauxite Residue: Roles of Extracellular Polymeric Substances Secreted by Penicillium oxalicum

Stable Aggregate Formation and Microbial Diversity Resilience in Soil Formation of Bauxite Residue: Roles of Extracellular Polymeric Substances Secreted by <i>Penicillium oxalicum</i>

Regulation and role of extracellular polymeric substances in the defensive responses of Dictyosphaerium sp. to enrofloxacin stress

Algae are susceptible to enrofloxacin (ENR), an antibiotic frequently detected in aquatic environments. However, algal responses, especially the secretion and roles of extracellular polymeric substances (EPS), under ENR exposure remain unknown. This study is the first to elucidate the variation in algal EPS triggered by ENR at both the physiological and molecular levels. The results showed that EPS were significantly (P < 0.05) overproduced along with increased polysaccharide and protein contents in algae exposed to 0.05, 0.5, and 5 mg/L ENR. Secretion of aromatic proteins, especially tryptophan-like substances with more functional groups or aromatic rings, was specifically stimulated. Furthermore, the genes with upregulated expression related to carbon fixation, aromatic protein biosynthesis, and carbohydrate metabolism are direct causes of enhanced EPS secretion. Improved EPS levels increased the cell surface hydrophobicity and provided more adsorption sites for ENR, which strengthened the van der Waals interaction and reduced ENR internalization. The hormesis effects of ENR were alleviated, as illustrated by the less affected cell density, chlorophyll a/b, and carotenoids biosynthesis in algae with EPS. These findings demonstrate the involvement of EPS in algal ENR resistance and promote a deeper understanding of the ecological effects of ENR in aquatic environments.

Biosorption of lanthanum by acidophile Euglena mutabilis biofilms and the role of extracellular polymeric substances

Enhancing the recovery of rare earth elements from wastewaters is of increasing interest given their growing economic and strategic importance as critical materials for a range of technologies including those required for renewable energy. We are particularly interested in the potential for using microalgal biofilms as sorbents given that they can grow as concentrated structures in the absence of organic carbon. In this study we assess and contrast biofilm and suspensions of Euglena mutabilis with respect to the kinetics and isotherms for the biosorption of lanthanum. The impact of extracellular polymeric substances (EPS) on biofilm biosorption was also examined. Suspension and biofilm adsorption kinetics were found to be similar with equilibrium being after approximately 2 to 3 h. Biofilms adsorbed significantly greater amounts of lanthanum compared to the suspension with a maximum adsorption capacity of 65 mg/g and 25 mg/g respectively for an increase of approximately 160 %. This difference in maximum adsorption capacities was attributed to the biofilm's increased EPS content which had an estimated adsorption capacity of 360 mg/g EPS. The proposed mechanism for EPS adsorption of lanthanum is via an ion exchange mechanism where calcium and magnesium were released into solution after lanthanum uptake. FTIR results also showed no change between the suspended Euglena and biofilm in terms of the available functional groups for adsorption.

Decoding the Role of Extracellular Polymeric Substances in Enhancing Nitrogen Removal from High-Ammonia and Low-C/N Wastewater in a Sequencing Batch Packed-Bed Biofilm Reactor

Although the role of extracellular polymeric substances (EPSs) as a viscous high-molecular polymer in biological wastewater treatment has been recognized, in-depth knowledge of how EPSs affect nitrogen removal remains limited in biofilm-based reactors. Herein, we explored EPS characteristics associated with nitrogen removal from high-ammonia (NH4+-N: 300 mg/L) and low carbon-to-nitrogen ratio (C/N: 2-3) wastewater in a sequencing batch packed-bed biofilm reactor (SBPBBR) under four different operating scenarios for a total of 112 cycles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and Fourier-transform infrared (FTIR) analysis revealed that the distinct physicochemical properties, interface microstructure, and chemical composition of the bio-carrier were conducive to biofilm formation and microbial immobilization and enrichment. Under the optimal conditions (C/N: 3, dissolved oxygen: 1.3 mg/L, and cycle time: 12 h), 88.9% ammonia removal efficiency (ARE) and 81.9% nitrogen removal efficiency (NRE) could be achieved in the SBPBBR. Based on visual and SEM observations of the bio-carriers, biofilm development, biomass concentration, and microbial morphology were closely linked with nitrogen removal performance. Moreover, FTIR and three-dimensional excitation-emission matrix (3D-EEM) spectroscopy demonstrated that tightly bound EPSs (TB-EPSs) play a more important role in maintaining the stability of the biofilm. Significant shifts in the number, intensity, and position of fluorescence peaks of EPSs determined different nitrogen removal. More importantly, the high presence of tryptophan proteins and humic acids might promote advanced nitrogen removal. These findings uncover intrinsic correlations between EPSs and nitrogen removal for better controlling and optimizing biofilm reactors.

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.

Performance Enhancement of Biogenetic Sulfidated Zero-Valent Iron for Trichloroethylene Degradation: Role of Extracellular Polymeric Substances.

Chemical sulfidation has been considered as an effective strategy to improve the reactivity of zero-valent iron (S-ZVI). However, sulfidation is a widespread biogeochemical process in nature, which inspired us to explore the biogenetic sulfidation of ZVI (BS-ZVI) with sulfate-reducing bacteria (SRB). BS-ZVI could degrade 96.3% of trichloroethylene (TCE) to acetylene, ethene, ethane, and dichloroethene, comparable to S-ZVI (97.0%) with the same S/Fe ratio (i.e., 0.1). However, S-ZVI (0.21 d-1) exhibited a faster degradation rate than BS-ZVI (0.17 d-1) based on pseudo-first-order kinetic fitting due to extracellular polymeric substances (EPSs) excreted from SRB. Organic components of EPSs, including polysaccharides, humic acid-like substances, and proteins in BS-ZVI, were detected with 3D-EEM spectroscopy and FT-IR analysis. The hemiacetal groups and redox-activated protein in EPS did not affect TCE degradation, while the acetylation degree of EPS increased with the concentration of ZVI and S/Fe, thus inhibiting the TCE degradation. A low concentration of HA-like substances attached to BS-ZVI materials promoted electron transport. However, EPS formed a protective layer on the surface of BS-ZVI materials, reducing its TCE reaction rate. Overall, this study showed a comparable performance enhancement of ZVI toward TCE degradation through biogenetic sulfidation and provided a new alternative method for the sulfidation of ZVI.

Pb(II) bioremediation using fresh algal-bacterial aerobic granular sludge and its underlying mechanisms highlighting the role of extracellular polymeric substances.

Lead (Pb) discharged from rural industries poses a significant threat to the environment and human health. Algal-bacterial aerobic granular sludge (A-B AGS) is a promising alternative for sewage treatment with high efficiency and good settleability. In this study, Pb(II) biosorption using fresh A-B AGS was investigated for the first time. The important role of extracellular polymeric substances (EPS) was revealed with the involved mechanisms being clarified. The desorbents for Pb recovery from Pb-loaded A-B AGS were also screened. Results showed that A-B AGS has an excellent maximum Pb adsorption capacity of 72.4mg·g-1 at pH 6.0. EPS plays an important role in keeping microbial activity, Pb bonding, and providing metal ions (Ca, Na and Mg) for Pb ion exchanges. Electrostatic interaction, ion exchange, and bonding to functional groups may occur orderly in the Pb biosorption process and the formation of pyromorphite (Pb5(PO4)3Cl) contributes to Pb biosorption. About 66% of the adsorbed Pb was accumulated in the A-B AGS microbial cells. Na2EDTA (0.05M) can recover 60.3% of the loaded Pb with the highest microbial activity of granules being remained. All the findings will provide the theoretical basis for the large-scale application of A-B AGS to bioremediate Pb(II)-containing wastewater.