•The current findings of the heterogeneous composition of flocs are summarized.•The effect of the composition of flocs on their structure and dynamics is reviewed.•The interaction among heterogeneous ...components within flocs is explained.•Human activities affecting the heterogeneous composition of flocs are also summarized.
Flocculation is a key process for controlling the fate and transport of suspended particulate matter (SPM) in water environments and has received considerable attention in the field of water science (e.g., oceanography, limnology, and hydrology), remaining an active area of research. The research on flocculation has been conducted to elucidate the SPM dynamics and to diagnose various environmental issues. The flocculation, sedimentation, and transportation of SPM are closely linked to the compositional and structural properties of flocs. In fact, flocs are highly heterogeneous in terms of composition. However, the lack of comprehensive research on floc composition and structure has led to misconceptions regarding the temporal and spatial dynamics of SPM. This review summarizes the current understanding of the heterogeneous composition of flocs (e.g., minerals, organic matter, metals, microplastic, engineered nanoparticles) and its effect on their structure and on their fate and transport within aquatic environments. Furthermore, the effects of human activities (e.g., pollutant discharge, construction) on floc composition are discussed.
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Microbial exopolysaccharides (EPS) are an abundant and important group of compounds that can be secreted by bacteria, fungi and algae. The biotechnological production of these substances represents a ...faster alternative when compared to chemical and plant-derived production with the possibility of using industrial wastes as substrates, a feasible strategy after a comprehensive study of factors that may affect the synthesis by the chosen microorganism and desirable final product. Another possible difficulty could be the extraction and purification methods, a crucial part of the production of microbial polysaccharides, since different methods should be adopted. In this sense, this review aims to present the biotechnological production of microbial exopolysaccharides, exploring the production steps, optimization processes and current applications of these relevant bioproducts.
In this study, we investigated the adsorption of Cd(II) and the biosynthesis of CdS quantum dots (QDs) mediated by cells of sulfate-reducing bacteria before and after the removal of EPS to determine ...whether EPS or the cell wall plays a major role. Potentiometric titration revealed that the concentration of proton-active binding sites on cells with EPS (EPS-intact) was notably higher than that on cells without EPS (EPS-free) and that the sites were predominantly carboxyl, phosphoryl, hydroxyl, and amine groups. The protein content in EPS-intact cells was higher, and thus the Cd(II) adsorption capacity was stronger. The CdS QDs biosynthesized using EPS-intact possessed better properties, including uniform size distribution, good crystallinity, small particle size, high fluorescence, and strong antimicrobial activity, and the yields were significantly higher than those of EPS-free by a factor of about 1.5–3.7. Further studies revealed that alkaline amino acids in EPS play a major role and serve as templates in the biosynthesis of QDs, whereas they were rarely detected in the cell wall. This study emphasizes the important role of EPS in the bacterial binding of metals and efficient recycling of hazardous waste in water.
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•Successful preparation of EPS-intact and EPS-free cells.•EPS play a more dominant role in mediating the synthesis of CdS QDs than cell wall.•EPS-mediated synthesis of CdS QDs has stronger ability and higher yield.•Alkaline amino acids act as biological templates in the synthesis of CdS QDs.
Tomato (Solanum lycopersicum) is a model for climacteric fleshy fruit ripening studies. Tomato ripening is regulated by multiple transcription factors together with the plant hormone ethylene and ...their downstream effector genes. Transcription Factors APETALA2a (AP2a), NON-RIPENING (NOR) and FRUITFULL (FUL1/TDR4 and FUL2/MBP7) were reported as master regulators controlling tomato fruit ripening. Their proposed functions were derived from studies of the phenotype of spontaneous mutants or RNAi knock-down lines rather than, as it appears now, actual null mutants. To study TF function in tomato fruit ripening in more detail, we used CRISPR/Cas9-mediated mutagenesis to knock out the encoding genes, and phenotypes of these mutants are reported for the first time. While the earlier ripening, orange-ripe phenotype of ap2a mutants was confirmed, the nor null mutant exhibited a much milder phenotype than the spontaneous nor mutant. Additional analyses revealed that the severe phenotype in the spontaneous mutant is caused by a dominant-negative allele. Our approach also provides new insight into the independent and overlapping functions of FUL1 and FUL2. Single and combined null alleles of FUL1 and FUL2 illustrate that these two genes have partially redundant functions in fruit ripening, but also unveil an additional role for FUL2 in early fruit development.
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•New procedure for separation of EPS into the main components from one sample.•Analysis of the main components of EPS from only one sample.•Characterization of EPS for a deeper ...insight into the state of the cells.•100 % recovery rates of synthetic EPS containing polysaccharides, proteins and lipids.•Successful transfer to real EPS samples from different cultivations.
Phototrophic biofilms produce a matrix of extracellular polymeric substances (EPS), which holds the cells together and functions inter alia as nutrient storage and protection layer. EPS mainly consist of water, polysaccharides, proteins, lipids and nucleic acids as well as lysis and hydrolysis products which makes the composition very complex. Thus, rough simplifications are used and commonly one or at most two components of the EPS are examined. In this work a new procedure for separation and analysis of EPS in the main components (i) polysaccharides, (ii) proteins and (iii) lipids is presented with recovery rates of nearly 100 %. The method was established with synthetic EPS, which based on the composition of real EPS described in literature. Afterwards, the method was transferred to real EPS samples allowing a deeper insight in the composition of EPS from only one sample. The composition of EPS-extracts from Nostoc spec, cultivated under heterotrophic and mixotrophic batch and fed-batch conditions, was analysed during a cultivation period of 14 days. It was observed that mixotrophic cultivation led to higher amounts of carbohydrates, lipids and proteins than heterotrophic cultivation respectively, regardless of batch or fed-batch culture. While the amount of proteins in the EPS increased during the cultivation period, carbohydrates and lipids were dominant in the beginning and decreased afterwards.
Chemical cleaning is vital for the optimal operation of membrane systems. Membrane chemical cleaning protocols are often developed in the laboratory flow cells (e.g., Membrane Fouling Simulator ...(MFS)) using synthetic feed water (nutrient excess) and short experimental time of typically days. However, full-scale Reverse Osmosis (RO) membranes are usually fed with nutrient limited feed water (due to extensive pre-treatment) and operated for a long-time of typically years. These operational differences lead to significant differences in the efficiency of chemical Cleaning-In-Place (CIP) carried out on laboratory-scale and on full-scale RO systems. Therefore, we investigated the suitability of lab-scale CIP results for full-scale applications. A lab-scale flow cell (i.e., MFSs) and two full-scale RO modules were analysed to compare CIP efficiency in terms of water flux recovery and biofouling properties (biomass content, Extracellular Polymeric Substances (EPS) composition and EPS adherence) under typical lab-scale and full-scale conditions. We observed a significant difference between the CIP efficiency in lab-scale (~50%) and full-scale (9–20%) RO membranes. Typical biomass analysis such as Total Organic Carbon (TOC) and Adenosine triphosphate (ATP) measurements did not indicate any correlation to the observed trend in the CIP efficiency in the lab-scale and full-scale RO membranes. However, the biofilms formed in the lab-scale contains different EPS than the biofilms in the full-scale RO modules. The biofilms in the lab-scale MFS have polysaccharide-rich EPS (Protein/Polysaccharide ratio = 0.5) as opposed to biofilm developed in full-scale modules which contain protein-rich EPS (Protein/Polysaccharide ratio = 2.2). Moreover, EPS analysis indicates the EPS extracted from full-scale biofilms have a higher affinity and rigidity to the membrane surface compared to EPS from lab-scale biofilm. Thus, we propose that CIP protocols should be optimized in long-term experiments using the realistic feed water.
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•CIP efficiency in full-scale RO is lower than “typical” lab-scale MFS.•CIP efficiency are not correlated to biomass removal indicators (ATP and TOC).•Full-scale RO biofilms have protein-rich EPS while lab-scale EPS has sugar-rich EPS.•Biofilm from full-scale EPS has higher affinity to membrane than lab-scale EPS.•CIP protocols should be developed using real feed water and for longer experiment.
Engineering C₄ photosynthesis into C₃ crops could substantially increase their yield by alleviating photorespiratory losses. This objective is challenging because the C₄ pathway involves complex ...modifications to the biochemistry, cell biology, and anatomy of leaves. Forward genetics has provided limited insight into the mechanistic basis of these properties, and there have been no reports of significant quantitative intraspecific variation of C₄ attributes that would allow trait mapping. Here, we show that accessions of the C₄ species Gynandropsis gynandra collected from locations across Africa and Asia exhibit natural variation in key characteristics of C₄ photosynthesis. Variable traits include bundle sheath size and vein density, gas-exchange parameters, and carbon isotope discrimination associated with the C₄ state. The abundance of transcripts encoding core enzymes of the C₄ cycle also showed significant variation. Traits relating to water use showed more quantitative variation than those associated with carbon assimilation. We propose that variation in these traits likely adapted the hydraulic system for increased water use efficiency rather than improving carbon fixation, indicating that selection pressure may drive C₄ diversity in G. gynandra by modifying water use rather than photosynthesis. The accessions analyzed can be easily crossed and produce fertile offspring. Our findings, therefore, indicate that natural variation within this C₄ species is sufficiently large to allow genetic mapping of key C₄ traits and regulators.
Biofilm structure and functionality depend on extracellular polymeric substances (EPS), but analytical methods for EPS often lack specificity which limits progress of biofilm research.
EPS were ...extracted from aerobic granular sludge and analyzed with frequently applied colorimetric methods. The colorimetric methods were evaluated based on their applicability for EPS analysis. EPS fractions of interest were proteins, sugars, uronic acids and phenolic compounds. The applied methods (Lowry method, bicinchoninic acid assay, phenol sulfuric acid method, carbazole sulfuric acid method) were investigated in terms of their sensitivity towards the selected standard compound. Interference of compounds present in EPS with the colorimetric methods was further evaluated. All methods showed to be highly depending on the choice of standard compound and susceptible towards interference by compounds present in EPS.
This study shows that currently used colorimetric methods are not capable of accurately characterizing EPS. More advanced methods are needed to be able to draw conclusions about biofilm composition, structure and functionality.
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•Colorimetric EPS analysis is very sensitive towards the standard selection.•Analysis of single EPS compounds is prone to interference by other EPS compounds.•EPS quantification with colorimetric methods can easily give inaccurate results.•New approaches and methods are needed to accurately characterize EPS.
► We study EPS from wild type and exoY strains of bacterium Sinorhizobium meliloti. ► EPS extracted by centrifugation, EDTA and formaldehyde/NaOH were compared. ► Two major components of ...proteinaceous and polysaccharide nature were found. ► Extraction methods influenced TOC, TN and total protein content in EPS. ► Molar mass distributions also differed among treatments and bacterial strains.
The influence of the extraction methods on the composition, size diversity, molar mass and size distributions of exopolymeric substances (EPS) from the bacterium Sinorhizobium meliloti wild type (WT) and by the exoY strain deficient in exopolysaccharide production was investigated. EPS obtained by centrifugation, EDTA and formaldehyde/NaOH were compared. It was found that the extraction method influenced TOC, TN and total protein content in EPS from both strains. However, no difference between EDTA and formaldehyde/NaOH methods was observed for the exopolysaccharide components. Similar functional groups and fluorescence pattern were found in the EPS obtained by different methods; however their relative abundance was method dependent. The extraction method also affected the molar mass and size distribution, HP SEC diversity among different treatment and bacterial strains.
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Extracellular polymeric substances (EPS) released by bacteria play a crucial role in microbial aggregation during wastewater treatment. Various environmental factors including ...substrate, anaerobic and aerobic conditions might contribute to the formation and properties of EPS, and thereby affect the properties of microbial aggregation and sludge. To reveal the relationship between these environmental factors and EPS properties, the characteristics of EPS generated in four types of sludge were investigated, including anammox granular sludge (ANMX), aerobic granular sludge (AGS) fed with glucose as carbon source (AGS-GLUC), AGS fed with liquor wastewater as carbon source (AGS-LIQ), and flocculent sludge from a real sewage plant (FLOC). Results indicated a positive correlation of EPS contents with granulation of sludge. As the dominant content of EPS, an increased proteins (PN) concentration facilitated the formation of microbial aggregates and the granulation of sludge. Bacteria in anaerobic environment (i.e., ANMX) or in aerobic environment with easily biodegradable carbon sources (i.e., AGS-GLUC) would generate more PN and EPS; yet, these conditions restrained the generation of humic acid (HA). The sludge from the sewage treatment plant (i.e., FLOC) had the lowest EPS and PN content, but the highest HA, showing flocculent structure sludge. In addition, a higher protein/polysaccharide (PN/PS) ratio and a lower zeta potential of EPS were conductive to sludge granulation. The hydrophobicity of EPS confirmed via analysis of chemical structures by FTIR would improve the microbial aggregation.
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