Journal
MICROBIOLOGY-SGM
Volume 146, Issue -, Pages 2671-2678Publisher
MICROBIOLOGY SOC
DOI: 10.1099/00221287-146-10-2671
Keywords
intracellular pH; Streptomyces puistinaespiralis; epifluorescence microscopy; image analysis
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Intracellular ph (pH(i)) is an essential parameter in the regulation of intracellular processes. Thus, its measurement might provide clues regarding the physiological state of cells cultivated in vitro, pH(i) of the filamentous, pristinamycin-producing Streptomyces pristinaespiralis was determined by epifluorescence microscopy and image analysis using the pH-sensitive fluorescent probe BCECF-AM [2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, acetoxymethyl ester]. Staining cell culture samples (OD660 = 1) of S. pristinaespiralis with 20 muM BCECF at 28 degreesC for 30 min yielded a green/red fluorescence ratio (R-527/600) that correlated with the pH(i) of the cells for values ranging from 6.5 to 8.5. When S. pristinaespiralis was cultivated in pristinamycin-producing conditions (in batch mode, with a constant external ph of 6.8), the measured pH(i) varied between 6.3 and 8.7. In fact, pH(i) correlated with the excretion of pristinamycins and glucose consumption during the production process.
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