Efficient and rapid purification of lentil alpha-galactosidase by affinity precipitation with alginate.
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 Km and Vmax 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.
Celem
Evran Biçak
EB
Department of Biochemistry, Faculty of Science, Ege University, Bornova-izmir, Turkey.
Bolle
Sharon Sibel
SS
Onal
Seçil
S
D000464:Alginates / Q000737:chemistry*; D011232:Chemical Precipitation; D018525:Germination; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D027806:Lens Plant / Q000201:enzymology* / Q000254:growth & development; D012639:Seeds / Q000201:enzymology / Q000254:growth & development; D013696:Temperature; D013997:Time Factors; D000519:alpha-Galactosidase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D000519:alpha-Galactosidase