Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
641302 | Separation and Purification Technology | 2013 | 6 Pages |
Abstract
In this work, a papain inhibitor extracted from seeds of Adenanthera pavonina was covalently immobilized onto glutaraldehyde modified polyaniline (PANIG). The immobilization was very efficient, presenting 54.24% inhibitor retention in the following conditions: immobilization time of 30 min, at 4 °C, pH 7.0, inhibitor concentration of 20 mg and 5 mg of PANIG. The resultant material (PANIG-I) was tested as stationary phase for the cysteine proteases papain, ficin and bromelain purification through bioaffinity chromatography. A commercial preparation of papain was efficiently purified, resulting in a single band with 25 kDa. SDS-PAGE of purified bromelain showed a characteristic band around 28 kDa and this procedure resulted in 0.89-fold purification and 32.4% yield. The purification process was more effective for ficin reaching 2.6-fold purification and a single band of 25 kDa in the SDS-PAGE. These results showed that the stationary phase containing A. pavonina inhibitors immobilized onto PANIG is a very promising material for a single step purification of cysteine proteases through bioaffinity chromatography.
Related Topics
Physical Sciences and Engineering
Chemical Engineering
Filtration and Separation
Authors
Adriane G. Gamboa, Karla A. Batista, Flavio M. Lopes, Kátia F. Fernandes,