Purification and characterization of α-N-acetylgalactosaminidase from Clostridium perfringens

H. Y. Hsieh, M. Mitra, D. C. Wells, D. Smith

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

α-N-Acetylgalactosaminidase from Clostridium perfringens is an exoglycosldase that degrades the human blood type A epitope. A highly purified preparation of α-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-α-D-galactosamlne 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 °C for 12 months. ELISA experiments demonstrated activity against blood type A epitope.

Original languageEnglish (US)
Pages (from-to)91-97
Number of pages7
JournalIUBMB Life
Volume50
Issue number2
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

Keywords

  • Blood type A epitope
  • Clostridium perfringens
  • α-N-Acetylgalactosaminidase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

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