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Volume 146, Issue 10

Intracellular pH determination of pristinamycin-producing by image analysis Free

Abstract

Intracellular pH (pH) is an essential parameter in the regulation of intracellular processes. Thus, its measurement might provide clues regarding the physiological state of cells cultivated . pH of the filamentous, pristinamycin-producing 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 (OD=1) of with 20 μM BCECF at 28 °C for 30 min yielded a green/red fluorescence ratio ( ) that correlated with the pH of the cells for values ranging from 65 to 85. When was cultivated in pristinamycin-producing conditions (in batch mode, with a constant external pH of 68), the measured pH varied between 63 and 87. In fact, pH correlated with the excretion of pristinamycins and glucose consumption during the production process.

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): BCECF-AM, 2’,7’-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, acetoxymethyl ester , epifluorescence microscopy , image analysis , intracellular pH , pHi, intracellular pH , R527/600, green/red fluorescence ratio (527/600 nm) and Streptomyces pristinaespiralis
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2000-10-01
2024-04-27
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Intracellular pH determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis
Microbiology 146, 2671 (2000); https://doi.org/10.1099/00221287-146-10-2671
/content/journal/micro/10.1099/00221287-146-10-2671
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