Purification of alpha-galactosidase by affinity precipitation with alginate.
Affinity precipitation is a technique which is known for over 20 years, but has recently received more attention due to the development of new materials for its implementation. It is a relatively simple, convenient, and reproducible technique that results in high target molecule recovery at high specificity. We describe, here, an efficient and rapid purification procedure for Vicia faba alpha-galactosidase (EC 3.2.1.22) by using affinity precipitation with alginate. The enzyme was purified with 43% activity yield and 40-fold purification. SDS-PAGE of the purified enzyme showed a single band and a subunit weight of 44 kDa. The properties of the enzyme were also searched. The results showed that the general properties of the enzyme offer potential for use of this alpha-galactosidase in several production processes.
Celem
Evran Biçak
EB
Department of Biochemistry, Faculty of Science, Ege University, Bornova-Izmir, Turkey.
Onal
Seçil
S
D000464:Alginates / Q000737:chemistry*; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D018525:Germination; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D012639:Seeds / Q000201:enzymology; D031307:Vicia faba / Q000196:embryology; D000519:alpha-Galactosidase / Q000737:chemistry / Q000302:isolation & purification*
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D000519:alpha-Galactosidase