Phosphorylated Chemiluminescent Probe Based on Spiro-cyclobutyl-phenoxy-dioxetane for the In Situ Light-Activated Sensing of Alkaline Phosphatase

High Resolution Image Download MS PowerPoint Slide Alkaline phosphatase (ALP) plays important biological roles for many living species, including bacteria. Here, we developed an ALP probe ( CL-A1 ) using the dephosphorylation of an in situ light-activated chemiluminescent species based on a spiro-cyclobutane-substituted dioxetane scaffold. A sequential light-induced oxidation and ALP-mediated dephosphorylation of CL-A1 rapidly generated intense chemiluminescence (CL), which was stronger than that of the commercial CL substrates AMPPD and APS-5 tested in the presence of a signal enhancer. Analysis using Escherichia coli strains confirmed the applicability of CL-A1 for the sensitive detection of endogenous ALP activity using in situ light activation.