Intracellular ph determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis

被引:29
作者
Corvini, PFX
Gautier, H
Rondags, E
Vivier, H
Goergen, JL
Germain, P
机构
[1] Inst Natl Polytech Lorraine, CNRS, Lab Sci Genie Chim, F-54505 Vandoeuvre Les Nancy, France
[2] Inst Natl Polytech Lorraine, Lab Fermentat & Bioconvers Ind, F-54505 Vandoeuvre Les Nancy, France
来源
MICROBIOLOGY-SGM | 2000年 / 146卷
关键词
intracellular pH; Streptomyces puistinaespiralis; epifluorescence microscopy; image analysis;
D O I
10.1099/00221287-146-10-2671
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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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页码:2671 / 2678
页数:8
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