Catalase purification from rat liver with iron-chelated poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-glutamic acid) cryogel discs

Prep Biochem Biotechnol. 2016 Aug 17;46(6):602-9. doi: 10.1080/10826068.2015.1085400.

Abstract

In this study, iron-chelated poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-glutamic acid) (PHEMAGA/Fe(3+)) cryogel discs were prepared. The PHEMAGA/Fe(3+) cryogel discs were characterized by elemental analysis, scanning electron microscopy, Fourier transform infrared spectroscopy, swelling tests, and surface area measurements. The PHEMAGA/Fe(3+) cryogel discs had large pores ranging from 10 to 100 µm with a swelling degree of 9.36 g H2O/g cryogel. Effects of pH, temperature, initial catalase concentration, and flow rate on adsorption capacity of the PHEMAGA/Fe(3+) cryogel discs were investigated. Maximum catalase adsorption capacity (62.6 mg/g) was obtained at pH 7.0, 25°C, and 3 mg/ml initial catalase concentration. The PHEMAGA/Fe(3+) cryogel discs were also tested for the purification of catalase from rat liver. After tissue homogenization, purification of catalase was performed using the PHEMAGA/Fe(3+) cryogel discs and catalase was obtained with a yield of 54.34 and 16.67 purification fold.

Keywords: Catalase; IMAC; cryogel disc; enzyme; glutamic acid; purification.

MeSH terms

  • Adsorption
  • Animals
  • Catalase / chemistry
  • Catalase / isolation & purification*
  • Cryogels*
  • Glutamic Acid / analogs & derivatives*
  • Glutamic Acid / chemistry
  • Hydrogen-Ion Concentration
  • Iron Chelating Agents / chemistry*
  • Liver / enzymology*
  • Methacrylates / chemistry*
  • Microscopy, Electron, Scanning
  • Osmolar Concentration
  • Rats
  • Temperature

Substances

  • Cryogels
  • Iron Chelating Agents
  • Methacrylates
  • N-methacryloyl-glutamic acid
  • Glutamic Acid
  • hydroxyethyl methacrylate
  • Catalase