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Aims: This study attempts to determine which of the exopolymeric substances are involved in the adherence and aggregation of a Bacillus licheniformis biofilm., Methods and Results: The involvement of extracellular proteins and eDNA were particularly investigated using DNase and proteinase K treatment. The permeability of the biofilms increased fivefold after DNase I treatment. The quantification of the matrix components showed that, irrespective to the enzyme tested, eDNA and amyloid-like polymers were removed simultaneously. Size-exclusion chromatography analyses supported these observations and revealed the presence of associated nucleic acid and protein complexes in the biofilm lysates. These data suggest that some extracellular DNA and amyloid-like proteins were closely interlaced within the matrix. Finally, confocal laser scanning microscopy imaging gave supplementary clues about the 3D organization of the biofilms, confirming that eDNA and exoproteins were essentially layered under and around the bacterial cells, whereas the amyloid-like fractions were homogeneously distributed within the matrix., Conclusion: These results confirm that some DNA-amyloid complexes play a key role in the modulation of the mechanical resistance of B. licheniformis biofilms., Significance and Impact of the Study: The study highlights the need to consider the whole structure of biofilms and to target the interactions between matrix components. A better understanding of B. licheniformis biofilm physiology and the structural organization of the matrix will strengthen strategies of biofilm control., (© 2017 The Society for Applied Microbiology.)

Temperature-phased anaerobic digestion with a 50-70 °C pre-treatment is widely proposed for sludge. Here, such a sludge pre-treatment (65 °C) was studied against the physical, enzymatic and biodegradation processes. The soluble and particulate fractions were analysed in terms of biochemical composition and hydrolytic enzymatic activities. Two kinetics of organic matter solubilisation were observed: a rapid transfer of the weak-linked biopolymers to the water phase, including sugars, proteins or humic acid-like substances, to the water phase, followed by a slow and long-term solubilisation of proteins and humic acid-like substances. In addition, during the heat treatment a significant pool of thermostable hydrolytic enzymes including proteases, lipases and glucosidases remains active. Consequently, a global impact on organic matter was the transfer of the biodegradable chemical oxygen demand (COD) from the particulate to the soluble fraction as evaluated by the biological methane potential test. However, the total biodegradable COD content of the treated sludge remained constant. The heat process improves the bio-accessibility of the biodegradable molecules but doesn't increase the inherent sludge biodegradability, suggesting that the chemistry of the refractory proteins and humic acids seems to be the real limit to sludge digestion.

Extracellular Polymeric Substances (EPS) analysis was undertaken on three biofilms grown under different feeding conditions and offering diverging microbial activities and structural characteristics. EPS were extracted by a multi-method protocol including sonication, Tween and EDTA treatments and were characterized by size exclusion chromatography (SEC). Tween and sonication extracts presented higher EPS size diversity compared to EDTA extracts. EPS size diversity also increased with microbial functions within the biofilms and a specific 25-50 kDa cluster was identified only in extracts from biofilms presenting autotrophic activity. Another specific size cluster (180 kDa) occurred in Tween extracts provided from the mechanically stable biofilms. Such specific EPS appear as potential indicators for describing microbial and structural properties of biofilms. This study brings new elements for designing EPS fractionation and shows that size distribution analysis is an interesting tool to relate EPS diversity with macro-scale characteristics of biofilms., (© 2010 Elsevier Ltd. All rights reserved.)

This study aimed to characterize biofilms from the paper industry and evaluate the effectiveness of enzymatic treatments in reducing them. The extracellular polymeric substances (EPS) extracted from six industrial biofilms were studied. EPS were mainly proteins, the protein to polysaccharide ratio ranging from 1.3 to 8.6 depending on where the sampling point was situated in the paper making process. Eight hydrolytic enzymes were screened on a 24-h multi-species biofilm. The enzymes were tested at various concentrations and contact durations. Glycosidases and lipases were inefficient or only slightly efficient for biofilm reduction, while proteases were more efficient: after treatment for 24 h with pepsin, Alcalase® or Savinase®, the removal exceeded 80%. Savinase® appeared to be the most adequate for industrial conditions and was tested on an industrial biofilm sample. This enzyme led to a significant release of proteins from the EPS matrix, indicating its potential efficiency on an industrial scale. [ABSTRACT FROM PUBLISHER]

Highlights: [•] EPS distribution was studied in two biofilms showing different microbial populations. [•] Cohesive layers of the biofilms were evidenced by applying hydrodynamic shear. [•] EPS content did not relate to biofilm cohesive properties. [•] For the first time hydrophobic EPS in biofilms were characterized using HIC method. [•] The strength of hydrophobic interactions seemed to be related to microbial activity. [Copyright &y& Elsevier]