title: A proteomic view of cell physiology of Bacillus licheniformis.
abstract: The still ongoing sequencing of Bacillus licheniformis at the Göttingen Sequencing Laboratory provides the basis for proteome studies of the bacterium. By using two-dimensional (2-D) electrophoresis and protein identification by mass spectrometry, we were able to create master gels for B. licheniformis cells grown either in minimal medium or in complex medium containing about 300 and 180 entries, respectively. With the DECODON Delta 2D software we identified the most abundant protein spots on the gels, which were shown to perform mainly basic metabolic functions in the cell such as translation, amino acid metabolism, glycolysis, and tricarboxylic acid (TCA) cycle. Based on the master gels, we were able to study the regulation of metabolic pathways such as glycolysis and TCA cycle. In cells grown in the presence of glucose a significant increase of the amount of some glycolytic enzymes (TpiA, GapA, Pgk, Pgm, Eno, Pyk) and of the pyruvate dehydrogenase (PdhA-D) was found. At the same time, there is a strong repression of almost all TCA cycle enzymes and of the ATP synthase. Glucose also stimulates the acetate kinase (AckA) and the phosphotransacetylase (Pta) which are known to be involved in the overflow metabolism in B. subtilis. Furthermore, we began developing proteomic signatures for growth of B. licheniformis in complex medium. For this purpose, we compared the proteome pattern of exponentially growing cells with that of cells in different stages during stationary phase. The most obvious proteomic signature indicates that cells during stationary phase are subjected to a severe oxidative stress and a resulting protein stress. Furthermore, the level of many vegetative proteins is strongly reduced when the growth is arrested after entry into stationary phase. The data indicate that proteomics can be a valuable tool to describe the physiological state of B. licheniformis cell populations, e.g., of cells growing in a bioreactor.
authors.affiliation: Institut für Mikrobiologie, Ernst-Moritz-Arndt Universität, Greifswald, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Voigt
authors.forename: Thomas
authors.initials: T
authors.lastname: Schweder
authors.forename: Dörte
authors.initials: D
authors.lastname: Becher
authors.forename: Armin
authors.initials: A
authors.lastname: Ehrenreich
authors.forename: Gerhard
authors.initials: G
authors.lastname: Gottschalk
authors.forename: Jörg
authors.initials: J
authors.lastname: Feesche
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Maurer
authors.forename: Michael
authors.initials: M
authors.lastname: Hecker
mesh_terms: D001407:Bacillus / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D002468:Cell Physiological Phenomena; D003470:Culture Media; D003600:Cytosol / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D018384:Oxidative Stress; D020543:Proteome / Q000378:metabolism / Q000502:physiology*; D040901:Proteomics / Q000379:methods*; D012984:Software
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D020543:Proteome