title: Evaluation of EPS extraction protocols from anaerobic sludge for gel-based proteomic studies.
abstract: 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.
authors.affiliation: Núcleo de Pesquisa em Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Brasil.
authors.forename: J A
authors.initials: JA
authors.lastname: Zorel
authors.affiliation: Departamento de Química, Universidade Federal de Ouro Preto (UFOP), Brasil.
authors.forename: S F
authors.initials: SF
authors.lastname: Aquino
authors.affiliation: Núcleo de Pesquisa em Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Brasil.
authors.forename: A L
authors.initials: AL
authors.lastname: Sanson
authors.affiliation: Núcleo de Pesquisa em Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Brasil; Departamento de Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Campus Morro do Cruzeiro s/n Cep, Minas Gerais 35400-000, Brasil E-mail: silvana@iceb.ufop.br.
authors.forename: W
authors.initials: W
authors.lastname: Castro-Borges
authors.affiliation: Núcleo de Pesquisa em Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Brasil; Departamento de Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Campus Morro do Cruzeiro s/n Cep, Minas Gerais 35400-000, Brasil E-mail: silvana@iceb.ufop.br.
authors.forename: S Q
authors.initials: SQ
authors.lastname: Silva
mesh_terms: D000693:Anaerobiosis; D002241:Carbohydrates / Q000032:analysis*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D011108:Polymers / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012722:Sewage / Q000032:analysis; D053719:Tandem Mass Spectrometry
chemical_list: D002241:Carbohydrates; D011108:Polymers; D011506:Proteins; D012722:Sewage