Intracellular ph determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis
Intracellular ph determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis
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DOI：
10.1099/00221287-146-10-2671
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发表时间：
2000-10-01
期刊：
影响因子：
2.8
通讯作者：
Germain, P
中科院分区：
文献类型：
--
作者：
Corvini, PFX;Gautier, H;Germain, P
细胞内pH（pH（i））是调节细胞内过程的重要参数。因此，其测量可能提供有关体外培养细胞的生理状态的线索。通过落射荧光显微镜和图像分析，使用pH敏感的荧光探针BCECF-AM [2 '，7'-双（2-羧乙基）-5（6）-羧基荧光素，乙酰氧基甲酯]，测定丝状的、产生原始霉素的Streptomyces pristinaespiralis的pH（i）。对S. pristinaespiralis与20 μ M BCECF在28 ℃孵育30分钟产生绿色/红色荧光比率（R-527/600），其与细胞的pH（i）相关，值范围为6.5 - 8.5。当S.在产生普那霉素的条件下（以分批模式，外部pH恒定为6.8）培养pristinaespiralis，测得的pH（i）在6.3和8.7之间变化。事实上，pH（i）与生产过程中原始霉素的排泄和葡萄糖消耗相关。
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.