beta-N-Acetylglucosaminidase [EC 3.2.1.30] from Vibrio alginolyticus H-8 was purified by column chromatography on DEAE-Sepharose FF, phenyl-Sepharose HP, Superdex 200HR, and Mono Q. The molecular weight of the enzyme was estimated by SDS-gel electrophoresis (SDS-PAGE) to be 75 kDa. The pI was 4.6. The activity was inhibited by Ag+, Hg2+, iodoacetate, p-chloromercuribenzoate, and N-ethylmaleimide.