Characterization of endo-alpha-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 beta1, 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 acceptors, such as monosaccharides, disaccharides, and sugar alcohols. Using this transglycosylation activity of the endoglycosidase, it may be possible to synthesize neo-oligosaccharides.
Ashida
H
H
Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan. reed@kais.kyoto-u.ac.jp
Yamamoto
K
K
Murata
T
T
Usui
T
T
Kumagai
H
H
D001212:Asialoglycoproteins / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000032:analysis; D004795:Enzyme Stability; D060748:Fetuins; D006596:Hexosaminidases / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D000509:alpha-Fetoproteins / Q000378:metabolism; D048809:alpha-N-Acetylgalactosaminidase
D001212:Asialoglycoproteins; D001426:Bacterial Proteins; D004187:Disaccharides; D060748:Fetuins; D008670:Metals; D000509:alpha-Fetoproteins; C017029:asialofetuin; D006596:Hexosaminidases; D048809:alpha-N-Acetylgalactosaminidase