Purification and characterization of alpha-N-acetylgalactosaminidase from Clostridium perfringens.
alpha-N-Acetylgalactosaminidase from Clostridium perfringens is an exoglycosidase that degrades the human blood type A epitope. A highly purified preparation of alpha-N-acetylgalactosaminidase was obtained from C. perfringens by salt precipitation, gel filtration, ion-exchange chromatography, chromatofocusing, and high-pressure liquid chromatography. The final preparation was homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with a molecular mass of 72.1 kDa. The enzyme was highly selective for terminal N-acetyl-alpha-D-galactosamine residues. No other substantial glycosidase activities, specifically neuraminidase, were detected. The pH optimum of the enzyme was between 6.5 and 7.0, and activity was unaffected by ionic strength. No protease activity was detected and enzyme activity was stable at 4 degrees C for 12 months. ELISA experiments demonstrated activity against blood type A epitope.
Hsieh
H Y
HY
Department of Pathology, University of Missouri-Columbia, 65212, USA.
Mitra
M
M
Wells
D C
DC
Smith
D
D
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003016:Clostridium perfringens / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006596:Hexosaminidases / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007525:Isoelectric Focusing; D012492:Salts / Q000378:metabolism; D048809:alpha-N-Acetylgalactosaminidase
D000939:Epitopes; D007477:Ions; D012492:Salts; D006596:Hexosaminidases; C482722:NAGA protein, human; D048809:alpha-N-Acetylgalactosaminidase