Purification and characterization of Phaseolus vulgaris α-D-galactosidase isozymes

M. Dhar, M. Mitra, J. Hata, O. Butnariu, Daniel Smith

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A highly purified preparation of α-D-galactosidase [E.C. 3.2.1.22] isozymes was obtained from Phaseolus vulgaris (pinto bean) seeds by extraction, salt precipitation, ion exchange, and affinity chromatography. The final preparation was homogeneous by SDS-PAGE but revealed isozymes of relative mass of 38.3 and 39.6 kDa. The N-terminal sequence for both isozymes was identical, LANGLAKT (one letter code for amino acids). Relative native molecular mass was estimated at 149.3 kDa by Sephacryl S-200 chromatography. Activity was unaffected by ionic strength at high enzyme concentrations, and was specific for α-D-galactoside conjugates. No protease or haemagglutinin activity was detected, and activity was stable at 4°C. Studies with soluble oligosaccharides demonstrated high activity against the selected straight and branched-chain substrates. The enzyme was active against terminal α1-3 galactosyl residues on human and rabbit erythrocyte membranes. Because of its activity against membrane glycoconjugates, these isozymes may have potential utility for modifying membrane epitopes on native erythrocytes.

Original languageEnglish (US)
Pages (from-to)1055-1062
Number of pages8
JournalBiochemistry and Molecular Biology International
Volume34
Issue number5
StatePublished - 1994
Externally publishedYes

Fingerprint

Galactosidases
Phaseolus
Isoenzymes
Purification
Membranes
Affinity chromatography
Galactosides
Glycoconjugates
Ion Exchange Chromatography
Hemagglutinins
Erythrocyte Membrane
Molecular mass
Enzymes
Chromatography
Ionic strength
Oligosaccharides
Affinity Chromatography
Osmolar Concentration
Seed
Epitopes

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Molecular Biology

Cite this

Purification and characterization of Phaseolus vulgaris α-D-galactosidase isozymes. / Dhar, M.; Mitra, M.; Hata, J.; Butnariu, O.; Smith, Daniel.

In: Biochemistry and Molecular Biology International, Vol. 34, No. 5, 1994, p. 1055-1062.

Research output: Contribution to journalArticle

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