A CTAB based method for the preparation of total protein extract of wine spoilage microrganisms for proteomic analysis.
Mapping the proteome of microrganisms by 2D-electrophoresis is often a hard task, because many contaminants, e.g. polysaccharides of the cell wall and nucleic acid, can obstruct the pores of the IEF gel resulting in streaks and smears. A protocol based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) and its salt-dependent solubility was developed. The cellulose-producing strain Gluconoacetobacter hansenii AAB0248 was resolved on 7cm Minigels in over 500 protein spots (a hundred more than with protocols reported in literature). The method was further employed for mapping the proteome of some acid adapted, wine spoilage microrganisms e.g. acetic acid bacteria and a yeast.
Polati
Rita
R
Dipartimento Scientifico e Tecnologico, Università di Verona, 37134 Verona, Italy.
Zapparoli
Giacomo
G
Giudici
Paolo
P
Bossi
Alessandra
A
D016947:Acetobacteraceae / Q000737:chemistry*; D001419:Bacteria / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D040901:Proteomics / Q000379:methods*; D014920:Wine / Q000032:analysis / Q000382:microbiology*; D015003:Yeasts / Q000737:chemistry
D001426:Bacterial Proteins; D002593:Cetrimonium Compounds; D000077286:Cetrimonium