Purification of Papain Using Reactive Green 5 Attached Supermacroporous Monolithic Cryogel

Uygun D. A. , Akduman B., Uygun M., AKGÖL S. , DENİZLİ A.

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, vol.167, no.3, pp.552-563, 2012 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 167 Issue: 3
  • Publication Date: 2012
  • Doi Number: 10.1007/s12010-012-9707-7
  • Page Numbers: pp.552-563


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.