Purification of α-Galactosidase by Affinity Precipitation with Alginate

Abstract

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 α-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 α-galactosidase in several production processes.

Keywords:

• Affinity macroligand
• Affinity precipitation
• Alginate
• Vicia faba
• α-Galactosidase

Notes

∗All experiments were performed in duplicate and the difference in the readings in the duplicates was less than ±5%.

Manuscript 7551