Comparison Between Fluorescent Probe and Ion-Selective Electrode Methods for Intracellular pH Determination in Leuconostoc mesenteroides

  • Curr Microbiol. 2018 Nov;75(11):1493-1497. doi: 10.1007/s00284-018-1550-9.
Duried Alwazeer  1 ,  Christophe Riondet  2 ,  Rémy Cachon  3
Affiliations
  • 1. Research Center for Redox Applications in Foods (RCRAF), Iğdır University, 76000, Iğdır, Turkey. [email protected].
  • 2. LGDP Laboratory of Genomes and Plant Development, Université de Perpignan, Perpignan, France.
  • 3. UMR Procédés Alimentaires et Microbiologiques A 02.102 AgroSup Dijon - Université de Bourgogne, 1 Esplanade Erasme, 21000, Dijon, France.
PMID: 30105525 DOI: 10.1007/s00284-018-1550-9
Abstract

The intracellular pH (pHin) of Leuconostoc mesenteroides subsp. mesenteroides 19D was evaluated by two different methods, fluorescent probe and ion-selective electrode. Two fluorescent probes 5 (and-6)-carboxyfluorescein diacetate succinimidyl ester (cFDASE) and 5 (and-6)-carboxy-2',7'-dichlorofluorescein diacetate succinimidyl ester (cDCFDASE) were tested to evaluate the intracellular pH (pHin) of living cells at a medium pH (pHex) ranged from 5.0 to 6.5. Salicylic acid was used as a probe for the ion-selective electrode method. Cells kept 60-80% of cFDASE probe at all pHex values against 5-10% of cDCFDASE probe at pHex ≤ 6.0. The pHin values measured by the ion-selective electrode were higher by 0.1-0.6 pH units at pHex ranged from 5.0 to 6.5 than those determinated by fluorescent probe method. The possibility to study the intracellular pH at a wide external pH range using a single probe, and the simplicity of the material and experimental protocol may make the ion-selective electrode method most useful and easy to measure the intracellular pH of lactic acid bacteria compared with the Other techniques like fluorescent probes.

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