Chemical Research in Chinese Universities ›› 2024, Vol. 40 ›› Issue (2): 287-292.doi: 10.1007/s40242-023-3255-2

• Articles • Previous Articles     Next Articles

Dynamic Measurement of Intracellular pH Based on Bioluminescent Bacteria

LI Yaohua, WANG Wei   

  1. State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, ChemBIC (Chemistry and Biomedicine Innovation Center), Nanjing University, Nanjing 210023, P. R. China
  • Received:2023-11-16 Revised:2023-12-07 Online:2024-04-01 Published:2024-03-27
  • Contact: WANG Wei wei.wang@nju.edu.cn
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (Nos. 21925403, 21874070), and the Excellent Research Program of Nanjing University, China (No. ZYJH004).

Abstract: Intracellular pH (pHi) is a fundamental indicator of cellular physiological state, regulating cellular state and function, and has important research values. Although various probes for measuring intracellular pH were available, it is challenging to reflect pHi in real-time and reversible manners. Herein, we developed a whole-cell bioluminescent (BL) probe based on wild type BL bacteria, Photobacterium phosphoreum (P. phosphoreum), to determine and image pHi. The dependence of BL intensity of P. phosphoreum on pH values of culture solutions was established. It was found that BL intensity could respond to the change of pH values rapidly and reversibly. We further revealed that P. phosphoreum maintained pH homeostasis in the extracellular pH (pHe) within the range of 5.0—7.0, while intracellular pH homeostasis was destroyed at the alkaline pHe. This method opens up the enormous potential of BL bacteria as an alternative to fluorescence for monitoring and imaging pHi.

Key words: Bioluminescence, P. phosphoreum, Intracellular pH, Cell imaging