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

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

Research output: Contribution to journalArticle

10 Citations (Scopus)

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
StatePublished - 2000
Externally publishedYes

Fingerprint

Clostridium
Clostridium perfringens
Purification
Epitopes
Blood
High pressure liquid chromatography
Glycoside Hydrolases
Neuraminidase
Enzyme activity
Molecular mass
Enzymes
Chromatography
Ionic strength
Electrophoresis
Sodium Dodecyl Sulfate
Ion exchange
Peptide Hydrolases
Salts
Gels
Ion Exchange Chromatography

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Purification and characterization of α-N-acetylgalactosaminidase from Clostridium perfringens. / Hsieh, H. Y.; Mitra, M.; Wells, D. C.; Smith, Daniel.

In: IUBMB Life, Vol. 50, No. 2, 2000, p. 91-97.

Research output: Contribution to journalArticle

Hsieh, H. Y. ; Mitra, M. ; Wells, D. C. ; Smith, Daniel. / Purification and characterization of α-N-acetylgalactosaminidase from Clostridium perfringens. In: IUBMB Life. 2000 ; Vol. 50, No. 2. pp. 91-97.
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