Purification and characterization of a Coffea canephora α-D-galactosidase isozyme

F. Haibach, J. Hata, M. Mitra, M. Dhar, M. Harmata, P. Sun, D. Smith

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Exoglycosidases modify carbohydrate epitopes on glycoproteins and glycolipids. The α-D-galactosidase from Coffea canephora is an important exoglycosidase which degrades the human blood group B epitope. Although multiple isozymes have been described, they have never been demonstrably purified and thoroughly characterized. We have developed a technique to purify an isozyme to homogeneity. The isolated enzyme has a molecular weight of 36.7 kDa by SDS PAGE and 34.0 kDa by gel filtration. The isozyme is highly selective for α-D-galactosides and inactive against other low molecular weight substrates. It hydrolyzes the the terminal α-D-galactosyl residue from the blood group B epitope. Protease activity is below detectable limits. The isozyme has a broad pH optima at 6.3, a pl of 7.03, is unaffected by ionic strength, and is stable at 4° C.

Original languageEnglish (US)
Pages (from-to)1564-1571
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume181
Issue number3
DOIs
StatePublished - Dec 31 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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