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Purification and partial characterization of β-glucosidase from papaya fruit

Authors
Journal
Phytochemistry
0031-9422
Publisher
Elsevier
Publication Date
Volume
25
Issue
10
Identifiers
DOI: 10.1016/s0031-9422(00)81677-5
Keywords
  • Carica Papaya
  • Caricaceae
  • Papaya Fruit
  • β-Glucosidase.
Disciplines
  • Biology

Abstract

Abstract β-Glucosidase (I) was isolated from Carica papaya fruit pulp and purified ca 1000-fold to electrophoretic homogeneity. The procedure used ammonium sulphate fractionation followed by chromatography on Phenyl-Sepharose CL-4B and Sephacryl S-200 to separate α-mannosidase (II) and, in part, β-galactosidase (III) from (I). Final separation of (III) from (I) was achieved by preparative isoelectric focusing (PIEF). The glycosidases had pI of 5.2 (I), 4.9 (II) and 6.9 (III). M,s of 54 000 (I), 260 000 (II) and 67 000 (III) were determined by gel filtration. The M, of (I) estimated by SDS-PAGE was 27 000 suggesting that (I) consisted of two subunits. The optimum pH and optimum temperature of (I) were 5.0 and 50°, respectively, and the enzyme followed typical Michaelis kinetics with K m and V max of 1.1 × 10 −4 M and 1.8 × 10 −6 mol/hr, respectively, for p-nitrophenyl-β- d-glucoside (40°).

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