Purification of papain using reactive green 5 attached supermacroporous monolithic cryogel.

Deniz Aktaş Uygun, Begüm Akduman, Murat Uygun, Sinan Akgöl, Adil Denizli

Chemistry Department, Faculty of Science and Arts, Adnan Menderes University, Aydin, Turkey. daktas@adu.edu.tr

Applied biochemistry and biotechnology 2012 Jun

filter terms:

Supermacroporous poly(2-hydroxyethyl methacrylate) [poly(HEMA)] monolithic cryogel was prepared by radical cryocopolymerization of HEMA with N,N'-methylene bisacrylamide as crosslinker. Reactive Green 5 dye was immobilized to the cryogel with nucleophilic substitution reaction, and this dye attached cryogel column was used for affinity purification of papain from Carica papaya latex. Reactive Green 5-immobilized poly(HEMA) cryogel was characterized by swelling studies, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy dispersive X-ray analysis. Maximum papain adsorption capacity was found to be 68.5 mg/g polymer while nonspecific papain adsorption onto plain cryogel was negligible (3.07 mg/g polymer). Papain from C. papaya was purified 42-fold in single step with dye attached cryogel, and purity of papain was shown by silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Citation

Deniz Aktaş Uygun, Begüm Akduman, Murat Uygun, Sinan Akgöl, Adil Denizli. Purification of papain using reactive green 5 attached supermacroporous monolithic cryogel. Applied biochemistry and biotechnology. 2012 Jun;167(3):552-63