A novel glycosphingolipid hydrolyzing enzyme, glycosphingolipid ceramide deacylase, which cleaves the linkage between the fatty acid and sphingosine base in glycosphingolipids.

Abstract

It has been demonstrated that the GM2 ganglioside cannot be cleaved by exo-beta-N-acetylhexosaminidases isolated from molds or bacterial sources. Here, a novel GM2 ganglioside-degrading enzyme was found in cells of Nocardia sp. This enzyme releases free fatty acids from the GM2 ganglioside. The chemical structure of the resultant lyso-GM2 ganglioside has been characterized by fast atom bombardment mass spectrometry, gas chromatography, and proton nuclear magnetic resonance spectroscopy. Using 14C-labeled GM2, at the fatty acid moiety, with stearic acid as the substrate, the optimum pH was determined to be 5.8. The enzyme was demonstrated to be capable of releasing fatty acids from GM3, GM2, GM1, and GD1a, and from neutral glycosphingolipids including Gb3-Cer, asialo-GM2, and asialo-GM1, but not from sphingolipids including Cer, Gal-Cer, Glc-Cer, and Lac-Cer. This enzyme, tentatively called glycosphingolipid ceramide deacylase, was found to be a tightly membrane-bound enzyme.