4.3 Article

Formation of volutin granules in Corynebacterium glutamicum

Journal

FEMS MICROBIOLOGY LETTERS
Volume 243, Issue 1, Pages 133-140

Publisher

WILEY-BLACKWELL PUBLISHING, INC
DOI: 10.1016/j.femsle.2004.11.047

Keywords

Corynebacterium glutamicum; inorganic polyphosphate; volutin granules; electron dispersive X-ray analysis; DAPI fluorescence; (31)P NMR; magnesium; polyphosphate glucokinase; MALDI-TOF; DNA microarray

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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) (31)P 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. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

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