Proteome analysis of Oenococcus oeni ATCC BAA-1163

PASCUALE RUSSO; VIVIAN DE LOS RIOS; MARÍA LAURA WERNING,; PILAR FERNÁNDEZ DE PALENCIA; PALOMA LÓPEZ; GIUSEPPE SPANO

Martina Franca

Simposio; 2º INTERNATIONAL SYMPOSIUM "MICRO SAFETY WINE”.; 2009

Abstarct Oenococcus oeni is an important lactic acid bacterium (LAB), which deacidifies wine, following the alcoholic fermentation, by means of the malolactic fermentation pathway. A better knowledge of the proteins involved in the metabolic pathways and transport systems of this bacterium would be of great utility to the wine industry. Therefore we report here on the characterization of the membranic, and the cytosolic proteomes of O. oeni ATCC BAA-1163, whose genomic DNA sequence is known. The preparation of proteins of subcellular fractions from LAB has been standardized. The O. oeni ATCC BAA-1163 membrane and cytosolic protein preparations were subjected to two-dimensional gel fractionation, by use of a non-linear pH gradient (pH 3.0-11.0) during electrofocusing. A comparative analysis of the proteomes from the two fractions was performed in order to establish whether the identified proteins were present in both or only one. Protein spots of interest were excised from the gel, digested with trypsin, and then analyzed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF/TOF). We identified 116 different polypeptides, which have been classified by their putative function and subjected to bioinformatics analysis to predict their subcellular location.