Results
Title

Purification of Papain Using Reactive Green 5 Attached Supermacroporous Monolithic Cryogel.

Authors

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

Abstract

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.

Subjects

POLYACRYLAMIDE gel electrophoresis; PAPAIN; ENERGY dispersive X-ray spectroscopy; FOURIER transform infrared spectroscopy; NUCLEOPHILIC substitution reactions; PAPAYA

Publication

Applied Biochemistry & Biotechnology, 2012, Vol 167, Issue 3, p552

ISSN

0273-2289

Publication type

Academic Journal

DOI

10.1007/s12010-012-9707-7