Efficient and rapid purification of lentil alpha-galactosidase by affinity precipitation with alginate

Celem E. B., BOLLE S. S., ÖNAL S.

INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS, vol.46, no.5, pp.366-370, 2009 (SCI-Expanded) identifier identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 46 Issue: 5
  • Publication Date: 2009
  • Journal Name: INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.366-370
  • Istanbul University Affiliated: No

Abstract

alpha-Galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) was purified (26-fold) from the germinating seeds of lentil (Lens culinaris) by affinity precipitation with alginate. The purified enzyme gave a single band corresponding to molecular-mass of 40 kDa on SDS-PAGE. The optimum temperature and pH of the enzyme were determined as 40 degrees C and 5.5, respectively. The enzyme was very stable at a temperature range of 4-65 degrees C and at a pH range of 4-7. The values of kinetic constants K-m and V-max using p-nitrophenyl-alpha-D-galactopyranoside (PNPG) as substrate were 0.191 mM and 0.73 U, respectively. Results suggest that affinity precipitation is an attractive process for the purification of alpha-galactosidase.