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 Citations (Scopus)

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

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Coffea
Galactosidases
Isoenzymes
Purification
Epitopes
Glycoside Hydrolases
Blood Group Antigens
Molecular Weight
Molecular weight
Galactosides
Glycolipids
Ionic strength
Osmolar Concentration
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Glycoproteins
Peptide Hydrolases
Gels
Carbohydrates
Substrates

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Purification and characterization of a Coffea canephora α-D-galactosidase isozyme. / Haibach, F.; Hata, J.; Mitra, M.; Dhar, M.; Harmata, M.; Sun, P.; Smith, D.

In: Biochemical and Biophysical Research Communications, Vol. 181, No. 3, 31.12.1991, p. 1564-1571.

Research output: Contribution to journalArticle

Haibach, F. ; Hata, J. ; Mitra, M. ; Dhar, M. ; Harmata, M. ; Sun, P. ; Smith, D. / Purification and characterization of a Coffea canephora α-D-galactosidase isozyme. In: Biochemical and Biophysical Research Communications. 1991 ; Vol. 181, No. 3. pp. 1564-1571.
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AU - Sun, P.

AU - Smith, D.

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