A series of water-soluble 2-(2'-arylsulfonamidophenyl)benzimidazole derivatives containing electron-donating and accepting groups attached to various positions of the fluorophore pi-system has been synthesized and characterized in aqueous solution at 0.1 M ionic strength. The measured pK(a)'s for deprotonation of the sulfonamide group of monosubstituted derivatives range between 6.75 and 9.33 and follow closely Hammett's free energy relationship. In neutral aqueous buffer, all compounds undergo efficient excited-state intramolecular proton transfer (ESIPT) to yield a strongly Stokes-shifted fluorescence emission from the phototautomer. Upon deprotonation of the sulfonamide nitrogen at high pH, ESIPT is interrupted to yield a new, blue-shifted emission band. The peak absorption and emission energies were strongly influenced by the nature of the substituents and their attachment positions on the fluorophore pi-system. The fluorescence quantum yield of the ESIPT tautomers revealed a significant correlation with the observed Stokes shifts. The study provides valuable information regarding substituent effects on the photophysical properties of this class of ESIPT fluorophores in an aqueous environment and may offer guidelines for designing emission ratiometric pH or metal-cation sensors for biological applications.