title: Protein extraction method for the proteomic study of Zymomonas mobilis during production of ethanol and levans.
abstract: Zymomonas mobilis has aroused considerable interest owing to its rapid metabolism and efficiency in producing ethanol and by-products such as levans, sorbitol, and gluconic acid from simple sugars. We performed a proteomic analysis of Z. mobilis UFPEDA241 to provide a global profile of regulatory proteins. The choice of the methods of extraction and cell lysis are fundamental steps and of great importance for the detection and identification of intra- and extracellular proteins of a proteome. Strains were subjected to protein extraction methods using three different reagents: TRIzol, lysis buffer, and phenol. The optimum method was taken to be the one that produced the greatest quantity and quality of proteins in one dimension for further analysis in two dimensions during the production of ethanol and levans over 72 h. The results showed that the greatest amount of protein was obtained by the phenol method (1.44 ± 0.07 mg/mL), which was significantly different (P < 0.05) to the TRIzol (1.11 ± 0.01 mg/mL), and lysis buffer (0.93 ± 0.01 mg/mL) methods (both with P > 0.05). Fermentation at 20°C produced the highest level of levans, and using two-dimensional electrophoresis and mass spectrometry it was possible to identify 34 differentially expressed spots.
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil danillocavalcanti@hotmail.com.
authors.forename: D R
authors.initials: DR
authors.lastname: Cavalcanti
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: C B
authors.initials: CB
authors.lastname: Malafaia
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: T D
authors.initials: TD
authors.lastname: Silva
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: B S
authors.initials: BS
authors.lastname: Santos
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: G M T
authors.initials: GM
authors.lastname: Calazans
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: M V
authors.initials: MV
authors.lastname: Silva
mesh_terms: D000431:Ethanol / Q000737:chemistry* / Q000378:metabolism; D005285:Fermentation; D005630:Fructans / Q000737:chemistry* / Q000378:metabolism; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D040901:Proteomics; D017355:Zymomonas / Q000235:genetics / Q000378:metabolism
chemical_list: D005630:Fructans; D011506:Proteins; D000431:Ethanol; D005947:Glucose