A New Chemiluminogenic Substrate for N-Acetyl-β-D-glucosaminidase, 4'-(6'-Diethylaminobenzofuranyl)phthalylhydrazido-N-acetyl-β-D-glucosaminide

Abstract

A new type of chemiluminogenic substrate for N-acetyl-β-D-glucosaminidase was synthesized. The substrate was obtained by incorporation of an enzyme-removable N-acetyl-D-glucosaminide group to the cyclic hydrazide moiety of an arylic hydrazide which contains a highly fluorescent benzofuran framework. Following enzyme-mediated hydrolysis, the substrate released the hydrazide that was subsequently subjected to chemiluminescent oxidation with hydrogen peroxide and a hemin catalyst in an alkaline buffer to generate light. The detection limit of the enzyme using this substrate was 0.6 I.U./I.