Formation of volutin granules in Corynebacterium glutamicum

• Published in: FEMS microbiology letters 2005-02, Vol.243 (1), p.133-140
• Main Authors: Pallerla, Srinivas Reddy, Knebel, Sandra, Polen, Tino, Klauth, Peter, Hollender, Juliane, Wendisch, Volker F., Schoberth, Siegfried M.
• Format: Article
• Language: English
• Subjects: 31P NMR; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biogenesis of cell structures, supramolecular organization; Biological and medical sciences; Cell size; Corynebacterium glutamicum; Corynebacterium glutamicum - genetics; Corynebacterium glutamicum - metabolism; Corynebacterium glutamicum - ultrastructure; Cytoplasmic Granules - metabolism; Cytoplasmic Granules - ultrastructure; DAPI fluorescence; DNA microarray; Electron dispersive X-ray analysis; Electron Probe Microanalysis; Ethylenediaminetetraacetic acids; Fluorescence; Fundamental and applied biological sciences. Psychology; Gel electrophoresis; Gene expression; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Glucokinase; Granular materials; Inorganic Chemicals - metabolism; Inorganic polyphosphate; Magnesium; Magnesium chloride; Magnesium Chloride - pharmacology; Magnetic Resonance Spectroscopy; MALDI-TOF; Microbiology; Microscopy, Electron, Transmission; NMR; Nuclear magnetic resonance; Oligonucleotide Array Sequence Analysis; Phosphorus; Phosphorus Isotopes; Phosphotransferases - metabolism; Polyphosphate glucokinase; Polyphosphates - metabolism; Proteins - metabolism; Sodium lauryl sulfate; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Staining; Transmission electron microscopy; Volutin granules
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1016/j.femsle.2004.11.047

Volutin granules are intracellular storages of complexed inorganic polyphosphate (poly P). Histochemical staining procedures differentiate between pathogenic corynebacteria such as Corynebacterum diphtheriae (containing volutin) and non-pathogenic species, such as C. glutamicum. Here we report that strains ATCC13032 and MH20-22B of the non-pathogenic C. glutamicum also formed subcellular entities (18–37% of the total cell volume) that had the typical characteristics of volutin granules: (i) volutin staining, (ii) green UV fluorescence when stained with 4′,6-diamidino-2-phenylindole, (iii) electron-dense and rich in phosphorus when determined with transmission electron microscopy and X-ray microanalysis, and (iv) 31P NMR poly P resonances of isolated granules dissolved in EDTA. MgCl 2 addition to the growth medium stimulated granule formation but did not effect expression of genes involved in poly P metabolism. Granular volutin fractions from lysed cells contained polyphosphate glucokinase as detected by SDS–PAGE/MALDI-TOF, indicating that this poly P metabolizing enzyme is present also in intact poly P granules. The results suggest that formation of volutin is a more widespread phenomenon than generally accepted.