Purification and properties of a β-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides

Sumathi Balasubramaniam, Heng Chin Lee, Hamid Lazan, Roohaida Othman, Zainon Mohd Ali

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

β-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. β-galactosidase I, II, III and IV. This β-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. β-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant β-galactosidases, thus, explaining the observed low similarity between the two subunits. β-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified β-galactosidase I was substantially active in hydrolyzing (1 → 4)β-linked spruce and a mixture of (1 → 3)β- and (1 → 6)β-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.

Original languageEnglish
Pages (from-to)153-163
Number of pages11
JournalPhytochemistry
Volume66
Issue number2
DOIs
Publication statusPublished - Jan 2005

Fingerprint

Galactosidases
Averrhoa carambola
galactosidases
Fruits
Cell Wall
Purification
Polysaccharides
Fruit
polysaccharides
Cells
cell walls
fruits
Protein Isoforms
Molecular mass
Amino Acid Sequence
polypeptides
amino acid sequences
Galactans
Pectins
Gum Arabic

Keywords

  • β-Galactosidase
  • Averrhoa carambola L.
  • Cell wall modification
  • Enzyme purification and properties
  • Fruit
  • Oxalidaceae

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery

Cite this

Purification and properties of a β-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides. / Balasubramaniam, Sumathi; Lee, Heng Chin; Lazan, Hamid; Othman, Roohaida; Ali, Zainon Mohd.

In: Phytochemistry, Vol. 66, No. 2, 01.2005, p. 153-163.

Research output: Contribution to journalArticle

Balasubramaniam, Sumathi ; Lee, Heng Chin ; Lazan, Hamid ; Othman, Roohaida ; Ali, Zainon Mohd. / Purification and properties of a β-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides. In: Phytochemistry. 2005 ; Vol. 66, No. 2. pp. 153-163.
@article{377a8f550943454886de61c170ffe3d4,
title = "Purification and properties of a β-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides",
abstract = "β-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. β-galactosidase I, II, III and IV. This β-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. β-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant β-galactosidases, thus, explaining the observed low similarity between the two subunits. β-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified β-galactosidase I was substantially active in hydrolyzing (1 → 4)β-linked spruce and a mixture of (1 → 3)β- and (1 → 6)β-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.",
keywords = "β-Galactosidase, Averrhoa carambola L., Cell wall modification, Enzyme purification and properties, Fruit, Oxalidaceae",
author = "Sumathi Balasubramaniam and Lee, {Heng Chin} and Hamid Lazan and Roohaida Othman and Ali, {Zainon Mohd}",
year = "2005",
month = "1",
doi = "10.1016/j.phytochem.2004.11.005",
language = "English",
volume = "66",
pages = "153--163",
journal = "Phytochemistry",
issn = "0031-9422",
publisher = "Elsevier Limited",
number = "2",

}

TY - JOUR

T1 - Purification and properties of a β-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides

AU - Balasubramaniam, Sumathi

AU - Lee, Heng Chin

AU - Lazan, Hamid

AU - Othman, Roohaida

AU - Ali, Zainon Mohd

PY - 2005/1

Y1 - 2005/1

N2 - β-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. β-galactosidase I, II, III and IV. This β-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. β-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant β-galactosidases, thus, explaining the observed low similarity between the two subunits. β-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified β-galactosidase I was substantially active in hydrolyzing (1 → 4)β-linked spruce and a mixture of (1 → 3)β- and (1 → 6)β-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.

AB - β-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. β-galactosidase I, II, III and IV. This β-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. β-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant β-galactosidases, thus, explaining the observed low similarity between the two subunits. β-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified β-galactosidase I was substantially active in hydrolyzing (1 → 4)β-linked spruce and a mixture of (1 → 3)β- and (1 → 6)β-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.

KW - β-Galactosidase

KW - Averrhoa carambola L.

KW - Cell wall modification

KW - Enzyme purification and properties

KW - Fruit

KW - Oxalidaceae

UR - http://www.scopus.com/inward/record.url?scp=12344280142&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12344280142&partnerID=8YFLogxK

U2 - 10.1016/j.phytochem.2004.11.005

DO - 10.1016/j.phytochem.2004.11.005

M3 - Article

VL - 66

SP - 153

EP - 163

JO - Phytochemistry

JF - Phytochemistry

SN - 0031-9422

IS - 2

ER -