A new metal chelate affinity adsorbent for Cytochrome c


EMIR S., SAY R., Yavuz H., DENIZLI A.

BIOTECHNOLOGY PROGRESS, vol.20, no.1, pp.223-228, 2004 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 20 Issue: 1
  • Publication Date: 2004
  • Doi Number: 10.1021/bp03033t
  • Journal Name: BIOTECHNOLOGY PROGRESS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.223-228
  • Hacettepe University Affiliated: Yes

Abstract

We have prepared a novel metal-chelate adsorbent utilizing N-methaeryloyl-L-histidine methyl ester (MAH) as a metal-chelating ligand. MAH was synthesized by using methacryloyl chloride and L-histidine methyl ester dihydrochloride. Spherical beads with an average diameter of 75-125 mum were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAH carried out in an aqueous dispersion medium. Then, Cull ions were chelated directly on the chelating beads. Cu2+-chelated beads were used in the adsorption of cytochrome c (cyt c) from aqueous solutions. The maximum cyt c adsorption capacity of the Cu2+-chelated beads (658.2 mumol/g Cu2+ loading) was found to be 31.7 mg/g at pH 10 in phosphate buffer. The nonspecific cyt c adsorption on the naked PHEMA beads was 0.2 mg/g. Cyt c adsorption increased with increasing Cu2+ loading. Cyt c adsorption capacity was demonstrated for the buffer types with the effects in the order phosphate > HEPES > MOPS > MES > Tris-HCl. Cyt c molecules could be adsorbed and desorbed five times with these adsorbents without noticeable loss in their cyt c adsorption capacity.