Extracellular polymeric substance

Extracellular polymeric substances (EPSs) are natural polymers of high molecular weight secreted by microorganisms into their environment. EPSs establish the functional and structural integrity of biofilms, and are considered the fundamental component that determines the physicochemical properties of a biofilm. EPS in the matrix of biofilms provides compositional support and protection of microbial communities from the harsh environments. Components of EPS can be of different classes of polysaccharides, lipids, nucleic acids, proteins, lipopolysaccharides, and minerals. Components EPSs are mostly composed of polysaccharides (exopolysaccharides) and proteins, but include other macromolecules such as DNA, lipids and humic substances. EPSs are the construction material of bacterial settlements and either remain attached to the cell's outer surface, or are secreted into its growth medium. These compounds are important in biofilm formation and cells' attach

Co-culture of microalgae and bacteria for wastewater treatment

Thierry Mounir

To address the problem of insucient wastewater treatment and eutrophication of lakes and rivers, the symbiotic association of microalgae and bacteria has shown great potential and numerous advantages. First, co-cultures can exhibit ecient treatment of domestic wastewater, with the combination of high nutrients (nitrogen and phosphorus) removal abilities of the microalgae and ecient degradation of organic matter by bacteria. Second, the symbiosis between microalgae and bacteria exchanging CO2 and O2 makes aeration not necessary, thus signicantly reducing the operating costs of the wastewater treatment plant. Third, recovered biomass of algae could be used as highpro t products like biofuels and food source. Fourth, the xation of the atmospheric CO2 by the microalgae through photosynthesis would contribute to greenhouse gas reduction in the atmosphere. Finally, the decayed microalgal biomass could serve as organic carbon source for denitrication by bacteria. However, this technology is not well-known yet, and further research on the in uence of the operating parameters have to be conducted. Furthermore, its main drawback is the harvesting of the microalgae. Due to their small size, microalgae demonstrate a low settling rate of 3.6 x 10􀀀3 m/h. A solution is to enhance aggregation of the biomass through occulation, followed by granulation to reach fully-developed microalgal-bacterial consortia. These granules exhibit high treatment performances and high settleability. The rst objective of this project was to identify the optimal microalgae:bacteria ratio for COD and nutrients removal. The second objective was to form fully-developed granules, starting from a co-culture, and assess the in uence of dierent agitation speeds on the granulation. The photobioreactor with a microalgae:bacteria ratio of 75:25 expressed the most promising results in COD removal eciency (95% after 8 days), nitrogen removal eciency (94% after 9 days), phosphorus removal eciency (93% after 7 days), and biomass productivity (144 mg/L/d). In this photobioreactor, nitrogen was removed by a combination of assimilation into microalgal biomass and nitrication/denitrication by the bacteria. The COD was removed by degradation by bacteria and mixotrophic microalgae that used the organic matter as carbon source. Finally, phosphorus was removed by assimilation into microalgal biomass. Although obvious aggregation between microalgae and bacteria could be observed, no fully-developed granules were formed after 89 batches of 24 hours. Because of low COD and nitrogen removals, no starvation period was experienced by the co-cultures. Therefore, extracellular polymeric substances were not released in sucient amounts to enhance the granulation. Furthermore, excessive stirring (200 rpm) may have lead to microalgal cells damage, thus promoting bacterial growth over microalgal growth. On the other hand, slower agitation speeds (80 and 120 rpm) seemed to enhance microalgal growth. Overall, the agitation speed appeared to have an indirect impact on the treatment performances and the settleability of the co-cultures, by signicantly in uencing the microalgal and bacterial growths.

2020

Optimizing the extraction of extracellular DNA of aerobic granular biofilms

Katarzyna Bialek

Formation and properties of aerobic granules for enhanced biological phosphorus removal (EBPR) from sequencing batch reactor (SBR) strongly depend on the extracellular polymeric substances (EPS) in which the microorganisms are embedded. The composition of EPS varies between different species but mostly it is made up of proteins, polysaccharides and extracellular DNA (eDNA), that has been recently recognized as an important structural component. Although great amount of literature on the EPS extraction is available, till now, there is no standardized protocol for EPS extraction. Extraction methods involving multi step processes seem to be the most promising, although too harsh extraction procedures can lyse the cells and release intracellular compounds, which contaminate the EPS samples creating bias. Commonly employed intracellular markers of cell lysis i.e. Glucose-6-phosphate dehydrogenase (G6PDH) or 2-keto-3deoxyoctonate (KDO) are not always applicable, especially in systems where glycolysis does not occur and in mixed communities with gram-positive bacteria, respectively. Therefore a new method to measure cell lysis is needed. The activity of isocitrate dehydrogenase (ICDH), an ubiquitous enzyme found in all living organisms utilzing the Krebs (citric acid) cycle, has been assessed. Initial assays using this enzyme activity during EPS extraction revealed to be very sensitive to detect cell lysis in samples from mixed microbial communities as granular sludge biofilms cultivated in SBR. Preliminary results of EPS extraction using multi-step method and enzymatic extraction as well as selection of appropriate dyes to visualize and distinguish intracellular and extracellular DNA in granular samples from the reactor are discussed.

2016

Probing the growth and mechanical properties of Bacillus subtilis biofilms through genetic mutation strategies

Lei Wang, Chen Zhang

Bacterial communities form biofilms on various surfaces by synthesizing a cohesive and protective extracellular matrix, and these biofilms protect microorganisms against harsh environmental conditions. Bacillus subtilis is a widely used experimental species, and its biofilms are used as representative models of beneficial biofilms. Specifically, B. subtilis biofilms are known to be rich in extracellular polymeric substances (EPS) and other biopolymers such as DNA and proteins like the amyloid protein TasA and the hydrophobic protein BslA. These materials, which form an interconnected, cohesive, three-dimensional polymer network, provide the mechanical stability of biofilms and mediate their adherence to surfaces among other functional contributions. Here, we explored how genetically-encoded components specifically contribute to regulate the growth status, mechanical properties, and antibiotic resistance of B. subtilis biofilms, thereby establishing a solid empirical basis for understanding how various genetic engineering efforts are likely to affect the structure and function of biofilms. We noted discrete contributions to biofilm morphology, mechanical properties, and survival from major biofilm components such as EPS, TasA and BslA. For example, EPS plays an important role in maintaining the stability of the mechanical properties and the antibiotic resistance of biofilms, whereas BslA has a significant impact on the resolution that can be obtained for printing applications. This work provides a deeper understanding of the internal interactions of biofilm components through systematic genetic manipulations. It thus not only broadens the application prospects of beneficial biofilms, but also serves as the basis of future strategies for targeting and effectively removing harmful biofilms.

KEAI PUBLISHING LTD2022

Co-culture of microalgae and bacteria for wastewater treatment

Thierry Mounir

2020