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Purification and Characterization of a‐N‐Acetylgalactosaminidase from Clostridium perfringens
Author(s) -
Hsieh HsinYeh,
Mitra Moonmoon,
Wells Donald Charles,
Smith Daniel
Publication year - 2000
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/713803702
Subject(s) - clostridium perfringens , clostridium , characterization (materials science) , chemistry , microbiology and biotechnology , food science , biology , materials science , bacteria , nanotechnology , genetics
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.