title: Efficient recovery of whole cell proteins in Oenococcus oeni--a comparison of different extraction protocols for high-throughput malolactic starter applications.
abstract: In this study, we compared different total protein extraction protocols to achieve highly efficient isolation and purification of total proteins for the specific protein profiling of Oenococcus oeni. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns obtained for the different extraction protocols revealed not only a qualitative similar protein pattern but also quantitative variations with different intensity bands depending on the extraction method used. The selected extraction method added with sonication proved to work extremely well and efficiently and was able to obtain a high-resolution 2-D electrophoresis (2-DE) map. Prominent spots were successfully identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry and corresponded to 76 different proteins involved in the main metabolic pathways. The approach allowed to achieve a protein profiling specific for O. oeni from Aglianico wine with numerous characterized protein products corresponding to many different O. oeni genes and associated with main cellular pathways. Further investigations of the 2-DE protein expression profile will provide useful and interesting information on the molecular mechanisms at the protein level responsible for growth and survival of O. oeni in wine.
authors.affiliation: Dipartimento di Scienze, Università degli Studi della Basilicata, Viale dell'Ateneo Lucano 10, 85100, Potenza, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Cafaro
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Bonomo
authors.forename: Rocco
authors.initials: R
authors.lastname: Rossano
authors.forename: Marilena
authors.initials: M
authors.lastname: Larocca
authors.forename: Giovanni
authors.initials: G
authors.lastname: Salzano
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D008967:Molecular Biology / Q000379:methods*; D056585:Oenococcus / Q000737:chemistry* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014920:Wine / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins