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-trim...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2009-04, Vol.877 (10), p.887-891
Main Authors: Polati, Rita, Zapparoli, Giacomo, Giudici, Paolo, Bossi, Alessandra
Format: Article
Language:English
Subjects:
Acetic acid bacteria
Acetobacteraceae - chemistry
Analysis
Analytical, structural and metabolic biochemistry
Bacteria - chemistry
Bacterial Proteins - chemistry
Bacterial Proteins - isolation & purification
Biological and medical sciences
Cetrimonium Compounds - chemistry
Chemical Fractionation - methods
CTAB
Electrophoresis, Gel, Two-Dimensional
Fundamental and applied biological sciences. Psychology
General pharmacology
Medical sciences
Pharmacology. Drug treatments
Proteome
Proteomics - methods
Salt-dependent solubility
Vitaceae
Wine - analysis
Wine - microbiology
Wine spoilage
Yeast
Yeasts - chemistry
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Summary: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 7 cm 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.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2009.02.022