Efficient and rapid purification of lentil alpha-galactosidase by affinity precipitation with alginate
Makaleler > Tam Makale · 2009
Yazarlar: Evran Bicak Celem, Sharon Sibel Bolle, Seçil ÖNAL
INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS, SCI Expanded İndekslerine Giren Dergi, Uluslararası, 2009, cilt 46, 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.