PURIFICATION AND PROPERTIES OF 2 DEACETYLASES PRODUCED BY VIBRIO-ALGINOLYTICUS H-8

The Chitinase-producing bacterium Vibrio alginolyticus H-8 isolated fr om mud of Hamana Lake also produced two deacetylases for (GlcNAc)(2) e xtracellularly, Deacetylases DA1 and DA2 were purified from crude enzy me by column chromatography on Q-Sepharose FF, Phenyl Sepharose HP, Gi gapite, and Superdex 200 HR, The final preparation was homogeneous in SDS-PAGE, The molecular weights were 48,000 and 46,000 for deacetylase s DA1 and DA2, respectively, The pIs, optimum pHs, and optimum tempera tures for deacetylases DA1 and DA2 were as follows; DA1, pI 3.3, optim um pH 8.5-9.0, optimum temperature 45 degrees C, DA2, pI 3.5, optimum pH 8.0-8.5, optimum temperature 40 degrees C, Both deacetylases were s table at pHs between 7.0 and 11.0 and at temperatures below 40 degrees C, The activities of both enzymes were inhibited by Ag+ and Hg2+. H-1 -NMR of the reaction product by deacetylase DA1 for (GlcNAc)(2) showed that the purified deacetylase selectively hydrolyzed the 2-acetamide group at the reducing end of (GlcNAc)(2).