Extracellular polymeric substances diversity of biofilms grown under contrasted environmental conditions

Monique, Ras; Lefèbvre, Dominique; Derlon, Nicolas; Paul, Etienne; Girbal-Neuhauser, Elisabeth

doi:10.1016/j.watres.2010.11.021

Cited by 88 publications

(40 citation statements)

References 30 publications

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“…The biofilms were scraped and vigorously shaken and centrifuged. EPS were then extracted following the multi-methods sequential protocol previously described by Ras et al [35], but without sonication to prevent diatom lysis. One extraction sequence involving three steps was applied in sequence with intermediate centrifugations (3200g; 10 min) to collect the supernatants, which contained the solubilized EPS.…”

Section: Culture and Extraction Protocolsmentioning

confidence: 99%

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Double walled carbon nanotubes promote the overproduction of extracellular protein-like polymers in Nitzschia palea: An adhesive response for an adaptive issue

Verneuil

Silvestre

Randrianjatovo

et al. 2015

Carbon

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A B S T R A C TThe present study assessed the effects of double-wall carbon nanotubes (DWCNTs) dispersed in the presence of a realistic concentration of natural organic matter (NOM, 10 mg L À1 ) on the benthic diatom Nitzschia palea using toxicity tests and quantitative/qualitative extracellular polymeric substances (EPS) assays. No toxic effect was observed. A growth delay was measured after 48 h of exposure to concentrations of DWCNTs ranging from 1 mg L À1 ($29%) to 50 mg L À1 ($84%). Extracellular carbohydrates and proteins were extracted using a sequential multi-methods protocol to collect soluble, hydrophobic and ion-bridged extracellular polymeric substances (EPS). Extracted EPS were analyzed by colorimetric assays and size exclusion chromatography. The results highlighted a higher EPS concentration in exposed cultures that was primarily caused by an overproduction of protein-like polymers (protein or glycoproteins, PLPs). Such EPS overproduction and increase in proteins/carbohydrates ratio can partially explain the observed growth inhibition. EPS were preferentially extracted using hydrophobic conditions and were mainly composed of PLPs with either low (10 kDa) or high (174 kDa) molecular weights. These data highlights the affinity between DWCNTs and EPS, which is primarily driven by both physical and hydrophobic interactions. This indicates that N. palea can respond to DWCNTs by forming an EPS network optimized for adhering to and efficiently wrapping DWCNTs.

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Section: Culture and Extraction Protocolsmentioning

confidence: 99%

“…2), indicating that the amount of EPS produced by microorganisms can be driven by environmental conditions [35,48,49]. EPS allows for the adherence of benthic organisms to substrates.…”

Section: Dwcnt Effect On Eps Production and Possible Involvement In Gmentioning

confidence: 99%

Double walled carbon nanotubes promote the overproduction of extracellular protein-like polymers in Nitzschia palea: An adhesive response for an adaptive issue

Verneuil

Silvestre

Randrianjatovo

et al. 2015

Carbon

Self Cite

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“…Furthermore, there is no standard extraction procedure established, making difficult the meaning, comparison and interpretation of published results. However, recent studies tend to evaluate whether molecular diversity of EPS are potential markers for biofilm macro-scale characteristics [40]. …”

Section: Carbohydrate Content Of Exopolysaccharidesmentioning

confidence: 99%

Exopolysaccharides of the Biofilm Matrix: A Complex Biophysical World

Lembré¹,

Lorentz²,

Martino³

2012

The Complex World of Polysaccharides

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“…However, in vitro extraction and characterization of these amyloid proteins remain difficult due to their low solubility and aggregating properties. EPS extraction often constitutes the first step for ePN characterization, but the physical or chemical extraction procedures used differ widely, leading to a variety of EPS compositions for the extracts obtained (Ras et al 2011;Sheng et al 2010). Furthermore, quantification may be biased by the fact that it is not possible to extract all the biopolymers (Zuriaga-Agustí et al 2013) and also because some chemicals, such as EDTA, may interfere with the classic biochemical colorimetric methods for protein assay (Bourven et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Epicocconone, a sensitive and specific fluorescent dye for in situ quantification of extracellular proteins within bacterial biofilms

Randrianjatovo

Girbal-Neuhauser

Marcato-Romain

2015

Appl Microbiol Biotechnol

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Biofilms are ecosystems of closely associated bacteria encapsulated in an extracellular matrix mainly composed of polysaccharides and proteins. A novel approach was developed for in situ quantification of extracellular proteins (ePNs) in various bacterial biofilms using epicocconone, a natural, fluorescent compound that binds amine residues of proteins. Six commercial proteins were tested for their reaction with epicocconone, and bovine serum albumin (BSA) was selected for assay optimization. The optimized protocol, performed as a microassay, allowed protein amounts as low as 0.7 μg to as high as 50 μg per well to be detected. Addition of monosaccharides or polysaccharides (glucose, dextran or alginate) to the standard BSA solutions (0 to 250 μg ml(-1)) showed little or no sugar interference up to 2000 μg ml(-1), thus providing an assessment of the specificity of epicocconone for proteins. The optimized protocol was then applied to three different biofilms, and in situ quantification of ePN showed contrasted protein amounts of 22.1 ± 3.1, 38.3 ± 7.1 and 0.3 ± 0.1 μg equivalent BSA of proteins for 48-h biofilms of Pseudomonas aeruginosa, Bacillus licheniformis and Weissella confusa, respectively. Possible interference due to global matrix compounds on the in situ quantification of proteins was also investigated by applying the standard addition method (SAM). Low error percentages were obtained, indicating a correct quantification of both the ePN and the added proteins. For the first time, a specific and sensitive assay has been developed for in situ determination of ePN produced by bacterial cells. This advance should lead to an accurate, rapid tool for further protein labelling and microscopic observation of the extracellular matrix of biofilms.

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Extracellular polymeric substances diversity of biofilms grown under contrasted environmental conditions

Cited by 88 publications

References 30 publications

Double walled carbon nanotubes promote the overproduction of extracellular protein-like polymers in Nitzschia palea: An adhesive response for an adaptive issue

Double walled carbon nanotubes promote the overproduction of extracellular protein-like polymers in Nitzschia palea: An adhesive response for an adaptive issue

Exopolysaccharides of the Biofilm Matrix: A Complex Biophysical World

Epicocconone, a sensitive and specific fluorescent dye for in situ quantification of extracellular proteins within bacterial biofilms

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