Excited-state proton transfer (ESPT) in organic photoacids is a widely studied phenomenon in which D-luciferin is of special mention, considering the fact that apart from its phenolic OH group, the nitrogen atoms at either of the two thiazole moieties could also participate in hydrogen bonding interactions with a proton-donating solvent during ESPT. As a result, several transient species could appear during the ESPT process. We hereby deploy subpicosecond time-resolved fluorescence upconversion (FLUP) and transient absorption (TA) spectroscopic techniques to understand the detailed photophysics of D-luciferin in water as well as in dimethyl sulfoxide (DMSO) and ethanol. These transient-state spectroscopic studies reveal the population of two kinds of hydrogen-bonded (HB) complexes (HBCs) in the excited singlet (S1) state─HBC-I, which is formed at the OH site of the hydroxy benzothiazole moiety with a …