Biological origin of extracellular polymeric substance proteins in wastewater treatment plants Aalborg West, and Aalborg East: Metaproteomics
Author
Olesen, Michael Kruse
Term
10. term
Publication year
2012
Submitted on
2012-01-31
Pages
94
Abstract
Renseanlæg for spildevand (WWTP) har været undersøgt i årtier, blandt andet af Srinath m.fl. (1959), og er i dag veletablerede ingeniørsystemer. Alligevel er de biologiske processer, især samspillet mellem de mange mikroorganismer, endnu ikke fuldt forstået. Mere viden om de mikrobielle samfund (community ecology) kan forbedre styringen af anlæggene, hvilket er et hovedmål i EcoDesign-projektet. Nye metoder inden for genomik, transkriptomik og proteomik er blevet modne nok til at belyse komplekse systemer som WWTP. Fordi proteinudtryk afspejler mikrobiers aktivitet direkte, er proteiner afgørende for anlæggenes funktion og stabilitet. I dette studie undersøger vi proteinekstraktion og forsøger at identificere proteiner, der medvirker til biofilmdannelse. Biofilm er tynde, klæbrige lag af mikroorganismer, og deres matrix består af ekstracellulære polymere stoffer (EPS), herunder proteiner. Rå ekstrakter blev analyseret med SDS-PAGE (gelelektroforese) og LC-MS/MS (tandem massespektrometri). Ekstraktionsmetoderne viste sig ikke at være tilstrækkelige, hvilket førte til, at kun få proteiner blev identificeret. Til gengæld blev det humane enzym Elastase-3A fundet i alle anvendte metoder, hvilket giver yderligere indikation af, at dette resistente protein forekommer i renseanlæg. Rapporten samler den aktuelle viden og de opnåede resultater i dette felt og peger på behovet for forbedrede metoder samt videre undersøgelser.
Wastewater treatment plants (WWTPs) have been studied for decades, including by Srinath et al. (1959), and are now well-established engineering systems. However, the underlying biology—especially how diverse microorganisms interact—is not yet fully understood. Gaining insight into the community ecology of WWTPs can improve process control, which is a key goal of the EcoDesign project. Advances in genomics, transcriptomics, and proteomics now allow us to explore complex systems like WWTPs. Because protein expression directly reflects microbial activity, proteins are central to plant function and stability. In this study, we tested protein extraction methods and sought to identify proteins involved in biofilm formation. Biofilms are thin, sticky layers of microorganisms whose matrix is made of extracellular polymeric substances (EPS), including proteins. Crude extracts were analyzed using SDS-PAGE (gel electrophoresis) and LC-MS/MS (tandem mass spectrometry). The extraction methods were not sufficient, leading to only a few identified proteins. However, the human enzyme Elastase-3A was detected with all methods, adding evidence that this resistant protein is present in WWTPs. The report summarizes current knowledge and results in this area and highlights the need for improved methods and further studies.
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