Influence of extraction methods on proteins and carbohydrates analysis from MBR activated sludge flocs in view of improving EPS determination

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

“…Possible interference due to physical and/or chemical limitation of epicocconone binding was tested directly on the 48-h-old biofilms developed in a 96-well microplate by addition of BSA standard solutions. This assay was carried out according to the standard addition method (SAM) proposed by Zuriaga-Agustí et al (2013) for the quantification of proteins and carbohydrates in complex biological samples. Correct quantification of both secreted ePNs and added BSA was obtained for the three biofilms, indicating no significant limitations for in situ application of the epicocconone method up to 50 μg eq BSA per well.…”

“…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). Finally, in addition to being time-consuming, in vitro extraction of the EPS components leads to possible denaturation of ePN and to the alteration of their functional structure.…”

“…The sensitivity and specificity of the epicocconone assay were first established with soluble standard proteins before its implementation on biofilms. The quantification method was then applied to three different bacterial biofilms, allowing comparison of the amount of ePN secreted by each population after 48 h. Possible interferences of the matrix components on the analytical method were assessed using the previously described standard addition method (SAM) (Zuriaga-Agustí et al 2013). …”

Using adsorbent made from sewage sludge to enhance wastewater treatment and control fouling in a membrane bioreactor

“…EPS and SMP. Previous studies reported that EPS was the main cause of membrane fouling [35][36][37]. Lin et al [38] considered that EPS was the key biological material to significantly contribute to membrane fouling in MBRs.…”

Study on dyeing wastewater treatment at high temperature by MBBR and the thermotolerant mechanism based on its microbial analysis

“…The EPS extraction process was performed according to the protocol described [15], including soluble EPS, loosely bound EPS and tightly bound EPS. The protein and carbohydrates contents were determined referred to the previous reports [16].…”