Metal binding properties of extracellular polymeric substances extracted from anaerobic granular sludges

Extracellular polymeric substances (EPS) were extracted from four anaerobic granular sludges with different procedures to study their involvement in biosorption of metallic elements. EPS extracts are composed of closely associated organic and mineral fractions. The EPS macromolecules (proteins, polysaccharides, humic-like substances, nucleic, and uronic acids) have functional groups potentially available for the binding of metallic elements. The acidic constants of these ionizable groups are: pK _a1 (4–5) corresponding to the carboxyl groups; pK _a2 (6–7) corresponding to the phosphoric groups; pK _a3 (8–10) and pK _a4 (≈10) corresponding to the phenolic, hydroxyl, and amino groups. The polarographic study confirms the higher affinity of the EPS to bind to lead than to cadmium. Moreover, the binding of these metallic compounds with the EPS is a mix of several sorption mechanisms including surface complexation, ion exchange, and flocculation. Inorganic elements were found as ions linked to organic molecules or as solid particles. The mineral fraction affects the binding properties of the EPS, as the presence of salts decreases the EPS binding ability. Calcite and apatite particles observed on SEM images of EPS extracts can also sorb metallic elements through ion exchange or surface complexation.     

Comparison of the complexation potential of extracellular polymeric substances (EPS), extracted from activated sludges and produced by pure bacteria strains, for cadmium, lead and nickel

This paper provides information on the metal complexation potential of extracellular polymeric substances (EPS), extracted from activated sludges and from eight pure cultures of bacteria isolated from the same activated sludge. The EPS extracted from pure bacteria cultures are mainly composed of proteins and low quantities of polysaccharides and uronic acids in comparison with EPS extracted from activated sludges. The EPS studied present two apparent pK(a) and the IR spectra show the presence of the same functional groups on all the EPS studied. The ability of EPS to complex Cd, Pb and Ni, was studied at pH 7 with Chau and Ruzic's models using polarography titration. All of the EPS exhibited a greater ability to complex Pb than Ni, Cd showing the weakest affinity overall. The EPS extracted from the pure cultures of bacteria were less able to complex the metals than that extracted from activated sludges. Literature data, IR data and EPS phosphorous content, supported by the EPS pK(a), revealed that carboxylic and phosphoric groups may play a major role in binding to metals at pH 7. This study underlines the importance of metal exposure in order for bacteria to secrete or modify EPS. After exposure, the EPS then exhibit the greatest capacity to bind metal in order to protect bacteria from harmful effects of heavy metals.     

污泥脱水性能的关键影响因素研究

针对目前污泥脱水性能影响因素不明的问题,研究通过对SBR在7种运行工况条件下活性污泥胞外聚合物(extracellular polymeric substances,EPS)的提取和测试,并采用SPSS软件分析,探讨MLVSS/MLSS、胞外蛋白质/胞外多糖、胞外蛋白质含量、胞外多糖含量、EPS总含量对污泥脱水性能的影响。研究结果表明,MLVSS/MLSS、胞外蛋白质/胞外多糖是影响污泥脱水性能的关键因素,并建立了脱水污泥含水率和污泥结合水含量的影响因素模型:脱水污泥含水率=0.229×(MLVSS/MLSS)+0.023×(胞外蛋白质/胞外多糖)+0.539、污泥结合水含量=2.876×(MLVSS/MLSS)+0.335×(胞外蛋白质/胞外多糖)-0.012,通过对7个城市污水厂污泥脱水性能测试比较表明,模型对脱水污泥含水率和污泥结合水含量预测值相对误差分别在±1%和±5%以内,对污泥脱水性能预测效果良好。     

Tracking the spectroscopic and chromatographic changes of algal derived organic matter in a microbial fuel cell

Changes in the characteristics of algae-derived organic matter (AOM) were examined upon the operation of a microbial fuel cell (MFC) using multiple analytical methods. Temporal variations in the UV absorption and fluorescence excitation–emission matrix of the AOM revealed that less condensed humic-like components and large-sized protein-like fluorescent compounds were preferentially decomposed over the period of electricity generation. They also showed that low UV-absorbing extracellular organic matters (EOM) were produced at the end of the operation. SEC chromatograms demonstrated that smaller-sized UV-absorbing components were initially decomposed, followed by the net production of EOM with an intermediate molecular weight. Fourier transform infrared (FT-IR) spectra showed that proteins and polysaccharides were the two most dominant structures of the AOM in the MFC. Two-dimensional correlation spectroscopy combined with FT-IR provided additional valuable information on the sequential changes of the AOM, which occurred in the order of proteins → acidic functional groups → polysaccharides → amino acids/proteins.     

Quantification of the interactions between Ca,Hg and extracellular polymeric substances (EPS) of sludge

The interactions between metals (Ca²⁺ and Hg²⁺) and extracellular polymeric substances (EPS) extracted from the aerobic and anaerobic sludge in wastewater treatment reactors were investigated using a combination of zeta potential measurement and 3-dimensional excitation–emission matrix (EEM) fluorescence spectroscopy with parallel factor (PARAFAC) analysis. Results show that Ca²⁺ had no substantial effects on the EEM fluorescence spectra of the EPS, but their zeta potentials increased with the increasing Ca²⁺ dosage. However, Hg²⁺ had a significant effect on the EEM fluorescence spectra of the EPS, while their zeta potentials seemed not to be affected by the dose of Hg²⁺. The interactions between Hg²⁺ and EPS were elucidated using the fluorescence quenching with PARAFAC analysis, while the interactions between Ca²⁺ and EPS were evaluated by the zeta potential technique. The binding constants for Hg²⁺ and EPS were two orders of magnitude higher than those for Ca²⁺ and EPS, suggesting that the binding mechanisms between Ca²⁺ and EPS were different from those between Hg²⁺ and EPS. The results might be useful for understanding the roles of EPS in bacterial self-protection against heavy metals and the aggregate formation mechanisms through ionic bridging interactions.     

Copper (II) adsorption by the extracellular polymeric substance extracted from waste activated sludge after short-time aerobic digestion

The extracellular polymeric substance (EPS) extracted from waste activated sludge (WAS) after short-time aerobic digestion was investigated to be used as a novel biosorbent for Cu²⁺ removal from water. The EPS consisted of protein (52.6 %, w/w), polysaccharide (30.7 %, w/w), and nucleic acid (16.7 %, w/w). Short-time aerobic digestion process of WAS for about 4 h promoted the productivity growth of the EPS for about 10 %. With a molecular weight of about 1.9?×?10⁶ Da, the EPS showed a linear structure with long chains, and contained carboxyl, hydroxyl, and amino groups. The sorption kinetics was well fit for the pseudo-second-order model, and the maximum sorption capacity of the EPS (700.3 mg Cu²⁺/g EPS) was markedly greater than those of the reported biosorbents. Both Langmuir model and Freundlich model commendably described the sorption isotherm. The Gibbs free energy analysis of the adsorption showed that the sorption process was feasible and spontaneous. According to the complex results of multiple analytical techniques, including scanning electron microscopy, Fourier transform infrared spectroscopy, atomic force microscopy, etc., the adsorption process took place via both physical and chemical sorption, but the electrostatic interaction between sorption sites with the functional groups and Cu²⁺ is the major mechanism.     

Potential toxicity of amphenicol antibiotic: binding of chloramphenicol to human serum albumin

Antibiotics are widely used in daily life but their abuse has posed a potential threat to human health. To evaluate the toxicity of chloramphenicol (CAP) at the protein level, the interaction between CAP and human serum albumin (HSA) was investigated by fluorescence, Ultraviolet–visible (UV–Vis) absorption, Fourier transform infrared (FT-IR) spectroscopy and molecular docking methods. Fluorescence data revealed that the fluorescence quenching of HSA by CAP was the result of the formation of CAP–HSA complex, and the binding constant was determined to be 3.196?×?10⁴ L mol^?1 at 310 K. The thermodynamic determination indicated that the interaction was driven by enthalpy change and entropy change together, where the multiple hydrogen bonds (CAP and the residues Arg 222 and His 242 of HSA) and van der Waals forces were the dominant binding force. The site marker competition revealed that CAP bound into sub-domain IIA of HSA. The binding of CAP induced the drastic reduction in α-helix conformation and the significant enhancement in β-sheet conformation of HSA. Molecular docking study further confirmed the binding mode obtained by experimental study. This work provides a new quantitative evaluation method for antibiotics to cause the protein damage.     

Effect of NaCl on nitrification performance and extracellular polymeric substance characteristic of Klebsiella sp. TN-10

Klebsiella sp. TN-10, isolated from tannery wastewater, exhibited excellent nitrification ability cultivated in media with NaCl contents ranging from 0 to 35 g/L. Analysis of antioxidative enzyme activities of TN-10 stressed by NaCl showed that the stain produced more superoxide dismutase (SOD) and catalase (CAT) to fight against salt stress conditions. Moreover, the auto-aggregation index of TN-10 decreased with the increment of NaCl contents, which is consistent with results of zeta potentials. Extinction-emission matrix (EEM) and Fourier-transform infrared (FT-IR) spectrum analyses showed that the compositions of extracellular polymeric substances (EPS) by TN-10 after NaCl shock were similar. Furthermore, proteins and polysaccharides were the main components of EPS, and the contents were greatly influenced by salinity. Then, circular dichroism (CD) spectrum was used to calculate the contents of each protein secondary structure. These results provided further insights into the knowledge of nitrification characteristics, auto-aggregation performance, and EPS compositions of TN-10 during NaCl shock.

     

Lead and cadmium biosorption by extracellular polymeric substances (EPS) extracted from activated sludges: pH-sorption edge tests and mathematical equilibrium modelling

The sorption of Cd and Pb by extracellular polymeric substances (EPS) extracted from activated sludges originated from wastewater treatment plants (WWTPs) or Lab-scale bioreactors was investigated as a function of pH. The study was carried out using a polarographic method in the SMDE (stripping mercury dropping electrode) mode which is suited to determine labile metals in solution containing soluble ligands such as EPS. The results obtained provide evidence of the presence of a pH-sorption/desorption edge for Cd and Pb by EPS. The use of Kurbatov’s model gives information on the mechanisms involved through the determination of “relative complexation constants” (operationally defined) and the number of protons exchanged. The use of this model demonstrates that proton exchange with metals is not the only mechanism involved in metal biosorption by EPS. Other mechanisms such as cation exchange with Ca or Mg, global electric field surrounding the ligand or micro-precipitation of metals could be involved in metal sorption by EPS. The position of the pH-sorption edge curves and the “relative complexation constants” show that Pb displays a greater affinity for EPS than Cd. The studied EPS have large differences regarding binding strength of Cd and Pb. These differences are not correlated with the organic parameters measured to characterize the EPS, however the mineral fraction of the EPS could be involved to a large extent in the sorption of metal.     

磺胺和四环素类抗生素对活性污泥性能的影响简

废水处理工艺中抗生素类污染物的存在可能会对生物处理过程产生长期而深远的影响,为探明此类污染物对废水生物处理主体活性污泥性能等方面的影响,采用间歇培养法研究了活性污泥法处理污水时,抗生素类污染物的存在对活性污泥性能如胞外聚合物（EPS）、污染物处理能力、脱氢酶活性和群落结构的影响。结果表明,抗生素的存在会导致活性污泥的胞外聚合物总量及其主要组分蛋白质和多糖增加,以产生保护屏障;且由于污泥絮体解体,细胞破裂导致EPS中DNA和色氨酸含量增加。同时,由于蛋白质大量增加引起的表面负电荷的增加,使污泥疏水性增强,絮凝性能恶化;污泥絮体解体导致污泥颗粒变小,SVI也随之下降;在活性污泥脱氢酶活性急剧下降的同时,出水TOC迅速升高。此外,抗生素类污染物在抑制活性污泥中大部分细菌的同时,对部分菌群也有刺激生长作用,最终导致活性污泥生物群落结构的改变。四环素类抗生素对活性污泥的EPS和絮凝沉降性能的影响大于磺胺类,而对污水处理能力和群落结构的影响则不如磺胺类。抗生素类污染物的长期存在会对活性污泥沉降性能、絮凝性能、脱氢酶活性以及活性污泥群落结构等产生一系列负面影响,进而影响污染物去除效果,导致出水水质恶化。     

Investigation on the interaction between triclosan and bovine serum albumin by spectroscopic methods

Abstract

Multi-spectroscopic and molecular docking methods were used to study the interaction between triclosan (TCS) and bovine serum albumin (BSA). The results indicated that the fluorescence quenching of BSA by TCS was due to the formation of TCS–BSA complex through static quenching. This result was also demonstrated by time-resolved fluorescence experiment. The binding constants and number of binding sites between TCS and BSA were 1.30?×?10⁵ M^?1 and 1.17 at 298?K, respectively. The thermodynamic parameters were studied in detail which suggested that hydrophobic forces and hydrogen bond played major roles in the TCS–BSA interaction. Moreover, the site marker competitive experiments and docking studies revealed that TCS could bind BSA into site I in subdomain IIA. All the results of UV–vis spectrophotometry, circular dichroism spectroscopy and synchronous fluorescence spectroscopy showed that interaction between TCS and BSA induced conformation changes of BSA.     

Influence of extracellular polymeric substances (EPS) on Cd adsorption by bacteria

The role of extracellular polymeric substances (EPS) in Cd adsorption by Bacillus subtilis and Pseudomonas putida was investigated using a combination of batch adsorption experiments, potentiometric titrations, Fourier transform infrared spectroscopy (FTIR). An increased adsorption capacity of Cd was observed for untreated bacteria relative to that for EPS-free bacteria. Surface complexation modeling of titration data showed the similar pK_a values of functional groups (carboxyl, phosphate and hydroxyl) between untreated and EPS-free bacteria. However, site concentrations on the untreated bacteria were found to be higher than those on the EPS-free bacteria. FTIR spectra also showed that no significant difference in peak positions was observed between untreated and EPS-free bacteria and carboxyl and phosphate groups were responsible for Cd adsorption on bacterial cells. The information obtained in this study is of fundamental significance for understanding the interaction mechanisms between heavy metals and biofilms in natural environments.     

Characterization of extracellular polymeric substances (EPS) from periphyton using liquid chromatography-organic carbon detection–organic nitrogen detection (LC-OCD-OND)

A protocol was developed to extract, fractionate, and quantitatively analyze periphyton extracellular polymeric substances (EPS), which obtains both information on the molecular weight (M _r) distribution and protein and polysaccharide content. The EPS were extracted from freshwater periphyton between July and December 2011. Organic carbon (OC) compounds from different EPS extracts were analyzed using liquid chromatography-organic carbon detection–organic nitrogen detection (LC-OCD-OND), and total protein and polysaccharide content were quantified. Four distinct OC fractions, on the basis of M _r, were identified in all extracts, corresponding to high M _r biopolymers (≥80–4 kDa), degradation products of humic substances (M _r not available), low M _r acids (10–0.7 kDa), and small amphiphilic/neutral compounds (3–0.5 kDa). Low C/N ratios (4.3?±?0.8) were calculated for the biopolymer fractions, which represented 16–38 % of the measured dissolved organic carbon (DOC), indicating a significant presence of high M _r proteins in the EPS. Protein and polysaccharide represented the two major components of EPS and, when combined, accounted for the measured DOC in extracts. Differences in specific OC fractions of EPS extracts over the course of the study could be quantified using this method. This study suggests that LC-OCD-OND is a new valuable tool in EPS characterization of periphyton.     

膜-生物反应器处理超市废水中污泥性质与膜污染关系分析

在以处理超市废水实际工程的基础上,通过对污泥性质中的胞外聚合物(EPS)、溶解性微生物产物(SMPs)、溶解性COD(SCOD)和污泥浓度(MLSS)变化的分析,得出EPS积累的原因是多方面的,而EPS的过度积累对沉降性能有一定的恶化作用;此外,EPS与SMPs有很好的相关性.作者采用SPSS软件对污泥性质与膜污染之间...     

Relation between extracellular polymers' composition and its ability to complex Cd,Cu and Pb

Activated sludges originated from wastewater treatment plants (WWTPs) play an important role in heavy metal removal from effluents. Extracellular polymers (ECP) form a major part of the activated sludge and are heavily involved in biosorption of heavy metals. The complexation of three heavy metals (Cd, Cu and Pb) with ECP extracted from six activated sludges, originated from different WWTPs, was investigated at pH 7.

ECP in the study were shown to be mainly composed of proteins, humic acids, uronic acids and polysaccharides along with smaller amounts of lipids and nucleic acids. IR spectra confirmed the presence of the functional groups usually found in the ECP and the content in each fraction was determined using colorimetric methods. The determination of surface charge was carried out on each ECP sample and allowed two pK_a values characteristic of two distinctive functional groups to be determined. At the pH used in the study, the value of these constants indicates that only one functional group is under protonated form.

A polarographic method was used to determine the complexation parameters (number of binding sites and complexation constant) of ECP solutions towards metals. The following orders were established for the number of binding sites: Cu > Pb  Cd and for the stability of the ECP–metal complex: Cd > Pb  Cu.

A matrix of correlation between the composition of the polymers and the complexation parameters showed that the number of binding sites and the complexation constant were strongly linked to proteins, polysaccharides and humic substances content.     

Novel insights into enhanced dewatering of waste activated sludge based on the durable and efficacious radical generating

This study aims to develop a high-efficiency radical oxidation process for enhancing the dewaterability of waste activated sludge (WAS). Radical scavenging studies combined with electron paramagnetic resonance (EPR) were carried out for the direct radical identification and effectiveness evaluation of radical oxidation. The results indicated that Fe(II)-activated CaO₂ can pose a superior effect on dewatering WAS due to its distinctive capacity of stable ?OH production and the high reaction efficiency of regulated-released ?OH with water-holding organics. The mechanism for the enhanced dewatering performance was also explored. The rupture of sludge colloidal flocs and the reduction of hydrophilic functional groups in loosely bound extracellular polymeric substances (LB-EPS) were found to be mainly responsible for the release of interstitial water and improved dewaterability, respectively. In addition, an inference about the relationship between interfacial water and zeta potential of different EPS fractions was established by the simultaneous measurement of the binding affinities of Ca²⁺ and Fe²⁺/Fe³⁺ for EPS and bound water content. All these results provide the direct evidence that Fe(II)-activated CaO₂ is a promising pretreatment reagent for sludge disposal.

Implications: Fe(II)-activated CaO₂ was first proposed to be highly effective in enhancing the dewaterability of waste activated sludge. Electron paramagnetic resonance (EPR) spectroscopy provided the direct evidence for the specific advantages of CaO₂, especially the capacity of durable and efficacious ?OH production leading to the excellent conditioning performance.     

Function of bacterial cells and their exuded extracellular polymeric substances (EPS) in virus removal by red soils

The potential influence of autochthonous microorganisms on virus fate in soil is usually determined through extreme conditions of sterilization vs. nonsterilization; however, the relative importance of microbial cells and their exudates remains unclear. In this study, bacterial cells (cell) were harvested, and their exuded extracellular polymeric substances (EPS) were extracted from three strains of bacteria, namely, Gram-negative bacteria Pseudomonas putida and Pseudomonas aeruginosa as well as Gram-positive bacterium Bacillus subtilis. This study aimed to evaluate virus removal in solutions in the presence of cell, EPS, and their combination (cell/EPS), as well as to investigate how their presence affects virus removal efficiencies by four red soils based on batch experiments. Results showed that virus removal percentage in solutions ranged from 11 to 23 in the presence of cells only and from 12 to 15 in the presence of EPS only. The removal percentage in the combined cell/EPS treatment can be estimated by summing the results achieved by the cell and EPS treatments, separately. Meanwhile, cell presence had a negligible effect on virus removal by red soils. EPS and combined cell/EPS significantly reduced virus removal by 20 to 69 % and 16 to 50 %, respectively, which indicated that EPS served a dominant function in reducing virus removal. This study clearly demonstrated that the prediction of virus removal by red soils must consider the effect of bacteria, especially those producing large quantities of EPS, which can be responsible for the underestimation of viral load in certain studies.     

胞外多聚物对活性污泥絮凝性能影响的研究进展

活性污泥的絮凝性能直接影响出水SS浓度,而胞外多聚物(EPS)是影响活性污泥絮凝性能的重要因素。从EPS的组分及其化学性质入手,结合现有的活性污泥絮凝机制评价了EPS在活性污泥絮凝性能中的重要作用,系统分析了EPS组分对污泥絮凝性能的影响,以及EPS产量和组分的影响因素,并就近年来的研究热点———松散层EPS、致密层EPS与活性污泥絮凝性能的关系进行了归纳,还对利用EPS提高活性污泥絮凝性能的技术应用进行了总结。最后,对该领域的研究方向进行了展望。     

Effect of humic acids on photodegradation of chloroacetanilide herbicides under UV irradiation

The effects of two humic acids (HAs) of different origins on the photodegradation of the chloroacetanilide herbicides acetochlor, propisochlor and butachlor were investigated in this study. One of the tested HAs was a standard sample that was purchased from a commercial source, and the other was isolated from the black soil of Northeast China. The photolysis of all three herbicides followed pseudo-first-order kinetics under ultraviolet (UV) irradiation conditions, regardless of whether HAs were present or not. Both HAs improved the photolysis rates of acetochlor in a dose-reversed way, whereas they inhibited butachlor degradation under all experimental concentrations. The two HAs differed in their effects on propisochlor photolysis, changing from enhancement to inhibition, depending on the origin and concentration of HAs. Element and Fourier Transform Infrared spectroscopy analyses showed that the isolated HAs had more polysaccharides and less aliphatic groups than the commercial HAs, and it was indicated that some characteristic radicals (C═O, O─H and phenolic hydroxyls) in HAs were involved in the photolysis of the herbicides. Gas chromatography/mass spectrometry (GC/MS) analyses indicated that the presence of HAs had no effects on the photolysis pathway and photoproduct species of the three herbicides.     

Effect of humic acids on photodegradation of chloroacetanilide herbicides under UV irradiation

The effects of two humic acids (HAs) of different origins on the photodegradation of the chloroacetanilide herbicides acetochlor, propisochlor and butachlor were investigated in this study. One of the tested HAs was a standard sample that was purchased from a commercial source, and the other was isolated from the black soil of Northeast China. The photolysis of all three herbicides followed pseudo-first-order kinetics under ultraviolet (UV) irradiation conditions, regardless of whether HAs were present or not. Both HAs improved the photolysis rates of acetochlor in a dose-reversed way, whereas they inhibited butachlor degradation under all experimental concentrations. The two HAs differed in their effects on propisochlor photolysis, changing from enhancement to inhibition, depending on the origin and concentration of HAs. Element and Fourier Transform Infrared spectroscopy analyses showed that the isolated HAs had more polysaccharides and less aliphatic groups than the commercial HAs, and it was indicated that some characteristic radicals (C=O, O-H and phenolic hydroxyls) in HAs were involved in the photolysis of the herbicides. Gas chromatography/mass spectrometry (GC/MS) analyses indicated that the presence of HAs had no effects on the photolysis pathway and photoproduct species of the three herbicides.