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, cilt.46, ss.366-370, 2009 (SCI İndekslerine Giren Dergi) identifier identifier identifier identifier

  • Cilt numarası: 46 Konu: 5
  • Basım Tarihi: 2009
  • Dergi Adı: INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS
  • Sayfa Sayıları: ss.366-370

Özet

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.