Accumulation of PHA granules in Cupriavidus necator as seen by confocal fluorescence microscopy

0464901 - ÚPT 2017 RIV NL eng J - Journal Article
Mravec, F. - Obruča, S. - Krzyžánek, Vladislav - Sedláček, P. - Hrubanová, Kamila - Samek, Ota - Kučera, D. - Benešová, P. - Nebesářová, Jana
Accumulation of PHA granules in Cupriavidus necator as seen by confocal fluorescence microscopy.
FEMS Microbiology Letters. Roč. 363, č. 10 (2016), fnw094:1-7. ISSN 0378-1097. E-ISSN 1574-6968
R&D Projects: GA ČR(CZ) GA15-20645S
Grant - others:GA MŠk(CZ) LO1211
Institutional support: RVO:68081731 ; RVO:60077344
Keywords : polyhydroxyalkanoates * intracellular granules * confocal fluorescence microscopy * bacterial cell diameters
Subject RIV: BH - Optics, Masers, Lasers; BH - Optics, Masers, Lasers (BC-A)
Impact factor: 1.765, year: 2016
http://femsle.oxfordjournals.org/content/363/10/fnw094

Many bacteria are capable of accumulating intracellular granules of polyhydroxyalkanoates (PHA). In this work, we developed confocal microscopy analysis of bacterial cells to study changes in the diameters of cells as well as PHA granules during growth and PHA accumulation in the bacterium Cupriavidus necator H16 (formerly Ralstonia eutropha). The cell envelope was stained by DiD (R) fluorescent probe and PHA granules by Nile Red. Signals from both probes were separated based on their spectral and fluorescence life-time properties. During growth and PHA accumulation, bacterial cells increased their length but the width of the cells remained constant. The volume fraction of PHA granules in cells increased during PHA accumulation, nevertheless, its value did not exceed 40 vol. % regardless of the PHA weight content. It seems that bacterial cultures lengthen the cells in order to control the PHA volume portion. However, since similar changes in cell length were also observed in a PHA non-accumulating mutant, it seems that there is no direct control mechanism, which regulates the prolongation of the cells with respect to PHA granules volume. It is more likely that PHA biosynthesis and the length of cells are influenced by the same external stimuli such as nutrient limitation.
Permanent Link: http://hdl.handle.net/11104/0263647