title: Purification of papain using reactive green 5 attached supermacroporous monolithic cryogel.
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.
authors.affiliation: Chemistry Department, Faculty of Science and Arts, Adnan Menderes University, Aydin, Turkey. daktas@adu.edu.tr
authors.forename: Deniz Aktaş
authors.initials: DA
authors.lastname: Uygun
authors.forename: Begüm
authors.initials: B
authors.lastname: Akduman
authors.forename: Murat
authors.initials: M
authors.lastname: Uygun
authors.forename: Sinan
authors.initials: S
authors.lastname: Akgöl
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000327:Adsorption; D002021:Buffers; D029441:Carica / Q000201:enzymology; D005591:Chemical Fractionation / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry*; D059985:Cryogels / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008086:Lissamine Green Dyes / Q000737:chemistry*; D010206:Papain / Q000737:chemistry* / Q000302:isolation & purification*; D011073:Polyamines / Q000737:chemistry*; D011102:Polyhydroxyethyl Methacrylate / Q000031:analogs & derivatives* / Q000737:chemistry; D016062:Porosity; D016622:Silver Staining; D014867:Water / Q000737:chemistry
chemical_list: D002021:Buffers; D004396:Coloring Agents; D059985:Cryogels; D008086:Lissamine Green Dyes; D011073:Polyamines; C056971:poly(2-hydroxyethyl methacrylate)-polyamine graft copolymer; D014867:Water; D011102:Polyhydroxyethyl Methacrylate; C024537:green 5; D010206:Papain