Purification and properties of neutral beta-N-acetylglucosaminidase from carp blood

A neutral beta-N-acetylglucosaminidase has been purified to homogeneity from carp blood by a seven-step procedure. It was localized in the cytosol of red blood cells. The purified enzyme was specific to beta-N-acetylglucosaminide and inactive to beta-N-acetylgalactosaminide. It was competitively inhibited by free N-acetylglucosamine, but not by free N-acetylgalactosamine. The optimum pH of the enzyme was 6.5, with a stable pH range of 7.0 to 11.0. The enzyme showed greater heat-lability and anodal electrophoretic mobility than acidic beta-N-acetylglucosaminidases. The Mr value, estimated by sucrose density gradient centrifugation, was 122,000, and the enzyme dissociated into two nonidentical subunits with Mr values of 66,000 and 53,000, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With respect to the major characteristics, the neutral enzyme in carp blood was supposed to be a counterpart of hexosaminidase C in human and other mammalian tissues.