Protein extraction from activated sludge: An analytical approach

Monique, Ras; Girbal-Neuhauser, Elisabeth; Paul, Etienne; Spérandio, Mathieu; Lefèbvre, Dominique

doi:10.1016/j.watres.2007.11.011

Cited by 125 publications

(35 citation statements)

References 44 publications

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“…Thus, the amount of proteins and humic-like substances cannot be taken into account for the EDTA extractions. Ras et al (2008) also have demonstrated that the colorimetric method used to determine the protein content can either overestimate or underestimate the protein concentrations in EPS samples. Thus, three assumptions may be hypothesized: (a) the measured protein content is the real one and differences between the samples really exist, (b) the chemicals used interfere with the protein determination and do not allow to know the real protein content, and (c) the colorimetric method badly estimates the protein content and the protein contents can be determined only by protocols that add no chemicals during the extraction.…”

Section: Organic Composition Of Eps Extracts Of Anaerobic Granular Slmentioning

confidence: 99%

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

d’Abzac

Bordas

Hullebusch

et al. 2009

Appl Microbiol Biotechnol

271

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The characteristics of the extracellular polymeric substances (EPS) extracted with nine different extraction protocols from four different types of anaerobic granular sludge were studied. The efficiency of four physical (sonication, heating, cationic exchange resin (CER), and CER associated with sonication) and four chemical (ethylenediaminetetraacetic acid, ethanol, formaldehyde combined with heating, or NaOH) EPS extraction methods was compared to a control extraction protocols (i.e., centrifugation). The nucleic acid content and the protein/ polysaccharide ratio of the EPS extracted show that the extraction does not induce abnormal cellular lysis. Chemical extraction protocols give the highest EPS extraction yields (calculated by the mass ratio between sludges and EPS dry weight (DW)). Infrared analyses as well as an extraction yield over 100% or organic carbon content over 1 gg −1 of DW revealed, nevertheless, a carry-over of the chemical extractants into the EPS extracts. The EPS of the anaerobic granular sludges investigated are predominantly composed of humic-like substances, proteins, and polysaccharides. The EPS content in each biochemical compound varies depending on the sludge type and extraction technique used. Some extraction techniques lead to a slightly preferential extraction of some EPS compounds, e.g., CER gives a higher protein yield.

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Section: Organic Composition Of Eps Extracts Of Anaerobic Granular Slmentioning

confidence: 99%

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

d’Abzac

Bordas

Hullebusch

et al. 2009

Appl Microbiol Biotechnol

271

View full text Add to dashboard Cite

show abstract

“…The main SMP components measured in this experiment were protein, polysaccharide, DNA, and humic acid. Protein and humic acid were measured with a modified Lowry method (Ras et al 2008), polysaccharide was measured with an anthrone colorimetric assay (Gaudy 1962), and DNA with a diphenylamine method (Brown and Lester 1980). Table 1 shows the water quality of the SBR effluents and the SMP organic composition.…”

Section: Smp In the Effluent Of Sbrmentioning

confidence: 99%

Influence of soluble microbial products (SMP) on wastewater disinfection byproducts: trihalomethanes and haloacetic acid species from the chlorination of SMP

Wei

Liu

Zhang

et al. 2010

Environ Sci Pollut Res

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“…Some researchers used the DNA content in extracted EPS to evaluate the damage to nuclei whereby high DNA content indicated contamination with nucleic materials Liu and Fang, 2002;Sheng et al, 2010). Some used the activity of the intracellular enzyme-glucose-6-phosphate dehydrogenase (G6PD) to evaluate the lysis of cell membranes (Wingender et al, 2001;Ras et al, 2008) while others used 2-keto-3-deoxyoctonate (KDO), a characteristic component of lipopolysaccharides in the cell wall of Gram-negative (G − ) bacteria, to evaluate the damage to cell walls (Kumada et al, 1993;Yu et al, 2011;Adav and Lee, 2008). However, the abovementioned tests for cellular materials have their limits: the content of DNA in EPS is uncertain and so the level of DNA contamination resulting from cellular damage is also uncertain; environmental conditions, such as high temperatures and acidic or alkaline pH, can deactivate G6PD (Wang et al, 2002), and only Gram-negative (G − ) bacteria contain KDO (Kumada et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge

Guo

Liu

Xiao

2014

Journal of Biotechnology

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a b s t r a c t Extracellular polymeric substances (EPS) are susceptible to contamination by intracellular substances released during the extraction of EPS owing to the damage caused to microbial cell structures. The damage to cell walls and cell membranes in nine EPS extraction processes of activated sludge was evaluated in this study. The extraction of EPS (including proteins, carbohydrates and DNA) was the highest using the NaOH extraction method and the lowest using formaldehyde extraction. All nine EPS extraction methods in this study resulted in cell wall and membrane damage. The damage to cell walls, evaluated by 2-keto-3-deoxyoctonate (KDO) and N-acetylglucosamine content changes in extracted EPS, was the most significant in the NaOH extraction process. Formaldehyde extraction showed a similar extent of damage to cell walls to those detected in the control method (centrifugation), while those in the formaldehydeNaOH and cation exchange resin extractions were slightly higher than those detected in the control. N-acetylglucosamine was more suitable than KDO for the evaluation of cell wall damage in the EPS extraction of activated sludge. The damage to cell membranes was characterized by two fluorochromes (propidium iodide and FITC Annexin V) with flow cytometry (FCM) measurement. The highest proportion of membrane-damaged cells was detected in NaOH extraction (26.54% of total cells) while membranedamaged cells comprised 8.19% of total cells in the control.

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Protein extraction from activated sludge: An analytical approach

Cited by 125 publications

References 44 publications

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

Influence of soluble microbial products (SMP) on wastewater disinfection byproducts: trihalomethanes and haloacetic acid species from the chlorination of SMP

Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge

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