Critical evaluation of heat extraction temperature on soluble microbial products (SMP) and extracellular polymeric substances (EPS) quantification in wastewater processes.

Abstract

While soluble microbial products (SMP) and extracellular polymeric substances (EPS) in wastewater bioprocesses have been widely studied, a lack of standard quantification procedures make it difficult to compare results between studies. This study investigated the effect of temperature on SMP and EPS profiles for biological nutrient removal (BNR) sludges and aerobic membrane bioreactor sludge by adapting the commonly used heat extraction and centrifugation scheme, followed by colorimetric quantification of the carbohydrate and protein fractions using the phenol-sulfuric acid (PS) and the bicinchoninic acid (BCA) methods, respectively. To overcome known inconsistencies in colorimetry, total carbon (TC), total nitrogen (TN), and fluorometry analyses were performed in tandem. SMP samples marginally benefitted from heat extraction, owing to their mostly soluble nature, while EPS profiles were greatly influenced by temperature. 60 °C appears to be a suitable general-purpose extraction temperature near the lysis threshold for the sludges tested. The PS method's misestimation due to lack of specificity was observed and contrasted by TC analyses, while the TN analyses corroborated the BCA assays. Fluorometry proved to be a sensitive and rapid analytical method that provided semi-quantitative information on SMP and EPS constituents, particularly its proteinaceous components, with positive implications for robust wastewater process control.