Characterization of endo-α-N-acetylgalactosaminidase from Bacillus sp and syntheses of neo-oligosaccharides using its transglycosylation activity

Endo-alpha-N-acetylgalactosaminidase was purified to homogeneity from the culture fluid of Bacillus sp. isolated from soil and characterized, The molecular mass of the enzyme was estimated as 110 kDa. The enzyme was stable at pH 4.0-10.0, up to 55 degrees C, and was most active at pH 5.0. The substrate specificity of the enzyme was strict for the disaccharide, galactosyl beta 1, 3 N-acetyl-D-galactosamine, bound to aglycone in alpha configuration. On the other hand, the specificity of the enzyme for the aglycone structure was fairly relaxed, The enzyme could transfer the disaccharide from para-nitrophenyl substrate to various accepters, such as monosaccharides, disaccharides, and sugar alcohols. Using this transglycosylation activity of the endoglycosidase, it may be possible to synthesize neo-oligosaccharides. (C) 2000 Academic Press.