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Within the frame of our study we investigated Microccocus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary zone electrophoresis (CZE). They form chains and clusters on a different scale, which can be reflected in the electropherograms. A low buffer concentration of Tris-borate and Na2EDTA containing a polymeric matrix of 0.0125% poly(ethylene) oxide (PEO) was used. Key factors were the standardization and optimization of CE conditions, buffer solution, and pretreatment of bacterial samples, which are not transferable to different bacterial strains, in general. The different compositions of the cell wall of on the one hand Gram-positive (M. luteus) and Gram-negative (N. cinerea) cocci and on the other hand Gram-negative, rod-shaped bacteria (P. fluorescens), are probably responsible for the different pretreatment conditions.
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The fluorescence excitation and emission spectra are reported for P-3 (bis-dehydro-B-carotene-2-carboxylic acid), the major carotenoid pigment of psychrotrophic M. roseus. The excitation spectrum and the absorption spectrum showed good agreement with respect to the position of their peak maxima. The study also demonstrates that P-3 binds to liposomes prepared from synthetic lipids (PC, DOPG, or CL) or the total lipids of a mutant colourless M. roseus. Binding of P-3 to the membranes was accompanied by a decrease in the fluorescence emission intensity and a blue shift in lambda(em) maximum by 15 to 20 nm. The quantum yield of P-3 was observed to be low (1.7 x 10(-5)).
Photoacoustic spectra of cells of M. roseus were recorded to study the in vivo characteristics and localisation of the carotenoid pigments in these cells. The PA spectra indicated that both the psychrotrophic and mesophilic strains had similar chromophores. The cis carotenoids were prominent in the psychrotrophic M.roseus whereas shorter polyenes were more prominent in mesophilic M.roseus. Further, depth profiling photoacoustic studies revealed that in both the strains of M.roseus the bulk of the chromophore was associated with the cell membrane.
Pigmentation in a psychrotrophic M.roseus was found to be increased when the bacteria were grown at 5 degrees C as compared to its pigmentation at 25 degrees C. In addition more polar pigments were synthesised at low temperature. The pigments were identified as bacterioruberins and were demonstrated to bind to synthetic membranes of phosphatidylcholine with almost equal affinity, irrespective of the polarity of the pigments.
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