scholarly journals Optimising extraction of extracellular polymeric substances (EPS) from benthic diatoms: comparison of the efficiency of six EPS extraction methods

2009 ◽  
Vol 60 (12) ◽  
pp. 1201 ◽  
Author(s):  
Eri Takahashi ◽  
Jérôme Ledauphin ◽  
Didier Goux ◽  
Francis Orvain
Keyword(s):  
Extracellular Polymeric Substances ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Benthic Diatoms ◽  
Universal Method ◽  
Intertidal Sediments ◽  
Electronic Microscopy ◽  
Glucose Content ◽  
Diatom Culture ◽  
High Yields

There is no universal method that can be applied to extract bound extracellular polymeric substances (EPS) from benthic diatoms of intertidal sediments without causing cell lysis. Six extraction methods were tested on a diatom culture of Navicula jeffreyi to establish the best compromise between high yields of carbohydrate extraction and minimum cell lysis. Extraction with distilled water provoked cell lysis (as already known). The five other extraction methods (dowex resin, artificial seawater of half salinity and extractions after pretreatment with gluteraldehyde by three methods: water, dowex water and dowex buffer) did not provoke cell lysis as shown by transmission electronic microscopy. This result was confirmed by the minimum release of internal compounds (protein, ATP) and by the low proportions of glucose in dowex-extracted EPS compared with the water-extracted EPS, from which the high glucose content must be inferred as contamination by the chrysolaminaran. The extraction with dowex resin resulted in the second-highest concentration of carbohydrate after the water extraction and the EPS were especially rich in deoxy sugars, hence increasing the hydrophobic feature of these substances. For these reasons, we recommend extraction with dowex, which is also the best method for extracting bound EPS from other biofilms such as in activated sludges.

scholarly journals Extraction of exopolymers from biofilms: the protective effect of glutaraldehyde

2003 ◽  
Vol 47 (5) ◽  
pp. 175-179 ◽  
Author(s):  
J. Azeredo ◽  
M. Henriques ◽  
S. Sillankorva ◽  
R. Oliveira
Keyword(s):  
Protective Effect ◽  
Pseudomonas Fluorescens ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Protective Agent ◽  
Universal Method ◽  
Alcaligenes Denitrificans ◽  
Pre Treatment ◽  
High Yields

The extraction of the exopolymeric matrix is a prerequisite to properly assessing the composition of the biofilm. Several extraction methods have already been developed, however, no universal method has yet been adopted because the compromise between high yields of extraction and minimum cell lysis is difficult to establish. In fact, most of the extraction methods promote leakage of intracellular material. The most common extraction methods, Dowex resin and sonication, were assayed in biofilms of Pseudomonas fluorescens and Alcaligenes denitrificans submitted to a pre-treatment with glutaraldehyde (GTA). The assessment of ATP released after extraction was used as a criterion of cell lysis. The results showed that GTA is a protective agent against cell lysis. The pre-treatment with GTA is particularly useful combined with sonication.

Biogenic Stabilization of Intertidal Sediments: The Importance of Extracellular Polymeric Substances Produced by Benthic Diatoms

2005 ◽  
Vol 49 (4) ◽  
pp. 501-512 ◽  
Author(s):  
J.F.C. de Brouwer ◽  
K. Wolfstein ◽  
G.K. Ruddy ◽  
T.E.R. Jones ◽  
L.J. Stal
Keyword(s):  
Extracellular Polymeric Substances ◽  
Benthic Diatoms ◽  
Intertidal Sediments

Comparison of extraction methods for quantifying extracellular polymers in biofilms

1999 ◽  
Vol 39 (7) ◽  
pp. 211-218 ◽  
Author(s):  
Xiaoqi Zhang ◽  
Paul L. Bishop ◽  
Brian K. Kinkle
Keyword(s):  
Extraction Method ◽  
Extracellular Polymeric Substances ◽  
Cell Lysis ◽  
Extraction Methods ◽  
The Other ◽  
Protein Yield ◽  
Total Carbohydrate ◽  
Extracellular Polymers ◽  
Sulfate Reducing ◽  
Edta Extraction

Five commonly used extraction methods - regular centrifugation, EDTA extraction, ultracentrifugation, steaming extraction and regular centrifugation with formaldehyde (RCF) - were selected to study their effectiveness and repeatability in extracting extracellular polymeric substances (EPS) from aerobic/sulfate reducing and nitrifying/denitrifying biofilm samples. Biofilm EPS extraction yields were represented by carbohydrate and protein concentrations; the amount of cell lysis during the extractions was indicated by DNA concentration. The results showed that analyzing wash waters is essential in quantifying biofilm EPS; the contribution of this step varied from 8-50% of the total carbohydrate yield, depending on the extraction method. Among the extraction methods, the RCF extraction gave the greatest carbohydrate yield, the steaming extraction gave the greatest protein yield, and the other three extraction methods gave approximately equivalent amounts of carbohydrate and proteins for both types of biofilm. DNA in the EPS was 27 times smaller than in the pellets, indicating no significant cell lysis occurred during the extractions.

Effective methods for extracting extracellular polymeric substances from Shewanella oneidensis MR-1

2016 ◽  
Vol 74 (12) ◽  
pp. 2987-2996 ◽  
Author(s):  
You-Fen Dai ◽  
Yong Xiao ◽  
En-Hua Zhang ◽  
Li-Dan Liu ◽  
Ling Qiu ◽  
...  
Keyword(s):  
Extracellular Polymeric Substances ◽  
Ethylenediaminetetraacetic Acid ◽  
Heating Temperature ◽  
Three Dimensional ◽  
Shewanella Oneidensis ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Bacterial Cells ◽  
Matrix Spectrum ◽  
Dry Cell

Extracellular polymeric substances (EPS) play crucial roles in bio-aggregate formation and survival of bacterial cells. To develop an effective but harmless method for EPS extraction from Shewanella oneidensis MR-1, five extraction methods, i.e. centrifugation (control), heating (40, 45, 50, and 60 °C), and treatments with H2SO4, ethylenediaminetetraacetic acid (EDTA) and NaOH, were examined, respectively. Results from scanning electron microscope and flow cytometric analyses indicate that MR-1 cells were severely broken by H2SO4, NaOH and heating temperature ≥45 °C. Proteins and polysaccharides in EPS extracted by heating at 40 °C were 7.12 and 1.60 mg g−1 dry cell, respectively. Although EDTA treatment had a relatively lower yield of EPS (proteins and polysaccharides yields of 5.15 and 1.30 mg g−1 dry cell, respectively), cell lysis was barely found after EPS extraction. Three peaks were identified from the three-dimensional excitation–emission matrix spectrum of each EPS sample, suggesting the presence of protein-like substances. Furthermore, the peak intensity was in good accordance with protein concentration measured by the chemical analysis. In short, heating (40 °C) and EDTA treatments were found the most suitable methods for EPS extraction considering the cell lysis and EPS content, composition and functional groups together.

Comparison of extraction methods for the characterization of extracellular polymeric substances from aggregates of three biofilm-forming phototrophic microorganisms

2018 ◽  
Vol 64 (11) ◽  
pp. 887-899 ◽  
Author(s):  
Emilie Loustau ◽  
Jean-Luc Rols ◽  
Joséphine Leflaive ◽  
Claire-Emmanuelle Marcato-Romain ◽  
Elisabeth Girbal-Neuhauser
Keyword(s):  
Cation Exchange ◽  
Extracellular Polymeric Substances ◽  
Exchange Resin ◽  
Cation Exchange Resin ◽  
Cell Lysis ◽  
Extraction Methods ◽  
Size Exclusion ◽  
Tween 20 ◽  
Nitzschia Palea ◽  
Phototrophic Microorganisms

This paper aims to define a robust procedure to extract extracellular polymeric substances (EPS) from aggregates of three benthic phototrophic microorganisms: the cyanobacterium Phormidium autumnale, the diatom Nitzschia palea, and the green alga Uronema confervicolum. This study focuses on the extraction efficiency of polysaccharide and protein EPS by using two physical methods (sonication, cation exchange resin) and three chemical methods (formamide, EDTA, Tween 20) with minimum cell lysis. Cell lysis was evaluated by monitoring chlorophyll a release. The results indicated that sonication or incubation of the algae aggregates with 0.25% Tween 20 induced a high level of cell lysis. A combined extraction approach, with an initial dispersing pretreatment (Ultra-Turrax, 13 500 r·min–1, 1 min), followed by formamide addition (0.22%) and then incubation with Dowex cation exchange resin (50 g per g of dry biomass), provided the highest amount of extracted EPS (mostly proteins), with low cell lysis. Furthermore, extracted EPS were characterized by size exclusion chromatography, and the obtained fingerprints revealed similar profiles for the three benthic microorganisms with a majority of low molecular weight polymers (400 to 11 300 Da). However, additional EPS of high (>600 000 Da) and intermediate (20 000 to 80 000 Da) molecular sizes were specifically detected in the diatom extracts.

scholarly journals Seasonal changes in the biochemical fate of carbon fixed by benthic diatoms in intertidal sediments

2017 ◽  
Vol 63 (2) ◽  
pp. 550-569 ◽  
Author(s):  
Tanja C. W. Moerdijk-Poortvliet ◽  
Peter van Breugel ◽  
Koen Sabbe ◽  
Olivier Beauchard ◽  
Lucas J. Stal ◽  
...  
Keyword(s):  
Seasonal Changes ◽  
Benthic Diatoms ◽  
Intertidal Sediments

Pengambilan senyawa polisakarida ekstraseluler dari mikroorganisme dalam lumpur aktif sebagai adsorben logam berat

2018 ◽  
Vol 5 (2) ◽  
pp. 453
Author(s):  
H Haryono ◽  
Tjandra Setiadi
Keyword(s):  
Heavy Metal ◽  
Activated Sludge ◽  
Extraction Method ◽  
Extracellular Polymeric Substances ◽  
Research Result ◽  
Freundlich Isotherm ◽  
Extraction Methods ◽  
Average Maximum ◽  
Heavy Metal Adsorbent ◽  
Main Component

Extracellular polysaccaride extraction from activated sludge microorganism as heavy-metal adsorbent. Microorganisms in the form of bioagregate are the main component of activated sludge. It generally has an ability to produce EPS (extracellular polymeric substances). The important components in EPS are polysaccharides and proteins. EPS recovery from the activated sludge may be done by many extraction methods. Six of the extraction methods to extract EPS from the activated sludge have been studied in this research, such are: Middle Speed Centrifugation, Regular Centrifugation, NaOH Extraction, EDTA Extraction, Steaming Extraction and Sonication. EPS solution in the following step would be tested its performances as adsorbent of copper heavy metal. The research result shows that the best extraction method for EPS extraction was NaOH Extraction method. The amount of the polysaccharides obtained was 18,09 mg EPS/g TSS. On other hand, Steaming Extraction was the most ineffective extraction method. This method gives the polysaccharides result in the least amount, i.e., about 4,96 mg EPS/g TSS. In this research, the protein content was not detected in the all used EPS solution. In the adsorption test, the adsorption phenomena of copper metal with EPS adsorbent fitted to the Freundlich isotherm adsorption equation. The values of Freundlich's a constant (k and n) each were 2,282 and 0,963. The average maximum adsmption capaciry of EPS to metal Cu was 88,34 mg Cu/gEPS.Keywords:  Adsorption,  Heavy  Metal,  Activated  Sludge,  Extraction Methods, Polysaccharides Extracellular, Copper.AbstrakMikroorganisme dalam bentuk bioagregat adalah merupakan penyusun utama lumpur aktif, memiliki sifat  umum  yaitu  dalam  kemampuannya memproduksi  SPE (Senyawa  Polimer  Ekstraselluler). Komponen  utama dari SPE adalah polisakarida  dan protein. Pengambilan SPE dari bioagregat dapat dilakukan dengan banyak metode ekstraksi.Pada penelitian ini telah dipelajari karakteristik enam  metode  ekstraksi, yaitu: Sentrifugasi Kecepatan Sedang, Sentrifugasi Reguler, Ekstraksi dengan NaOH. Ekstraksi dengan EDTA, Ekstraksi dengan Pemanasan dan Sonikasi. Larutan SPE yang diperoleh, kemudian diuji kinerjanya sebagai adsorben logam berat tembaga. Hasil penelitian menunjukkan bahwa  metode ekstraksi terbaik adalah metode Ekstraksi dengan NaOH, dengan perolehan polisakarida sebesar 18,09 mg/g TSS. Sedangkan Ekstraksi dengan pemanasan merupakan metode ekstraksi yang paling tidak efektif dengan perolehan polisakarida paling sedikit, yaitu sekitar 4,96 mg/g TSS. Pada penelitian ini tidak dijumpai adanya protein di dalam larutan SPE dari semua  metode ekstraksi yang diterapkan. Pada uji adsorpsi, peristiwa adsorpsi logam  Cu dengan larutan SPE lebih mengikuti persamaan adsorpsi isoterm Freundlich dengan harga konstanta k dan n masing-masing sebesar 2,282 dan 0,963. Kapasitas adsorpsi maksimum rata-rata SPE terhadap logam Cu adalah sebesar 88,34 mg Cu/g SPE.Kata kunci:  Adsorpsi, Logam Berat, Lumpur Aktif, Metode Ekstraksi, Polisakarida Ekstraselluler, Tembaga.

Evaluation of EPS extraction protocols from anaerobic sludge for gel-based proteomic studies

2015 ◽  
Vol 72 (4) ◽  
pp. 535-542 ◽  
Author(s):  
J. A. Zorel ◽  
S. F. Aquino ◽  
A. L. Sanson ◽  
W. Castro-Borges ◽  
S. Q. Silva
Keyword(s):  
Extracellular Polymeric Substances ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Background Level ◽  
Molecular Size ◽  
Extraction Methods ◽  
Extraction Yield ◽  
Size Exclusion ◽  
Anaerobic Sludge ◽  
Distribution Profile

Despite the importance of anaerobic sludge extracellular polymeric substances (EPSs), their characterization is limited to information regarding their chemical classes and molecular size. This work explores the possibility of using proteomic techniques to study the proteins present in this matrix. Thus, this paper compares eight EPS extraction methods regarding extraction yield, protein/carbohydrate ratio, size distribution profile and suitability to sodium dodecyl sulfate–polyacrylamide gel electrophoresis analyses. Despite the differences found in quantification and size exclusion chromatography assays, the band profile found for all methods was very similar. Considering the band pattern, extraction time and background level, heating method followed by ammonium sulfate precipitation proved to be the most appropriate method for gel-based analyses of anaerobic sludge EPS proteins.

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