Purification of Superoxide Dismutase with Chitosan Coated Metal Affinity Chromatography

摘要

A new and an inexpensive adsorbent with chitosan coated for immobilized metal affinity chromatography (IMAC) was studied . After double coating , the chitosan coated on silica beads could be up to 53.4 mg / g silica beads. When pH > 3.8 , the metal ligand Cu~(2+) was chelated on the coated chitosan with a bound capacity of 14.6 mg /g chitosan without introducing iminodiacetic acid(IDA). The chitosan coated adsorbent could be used in the purification of superoxide dismutase (SOD) from the supernatant of Saccaromyces cerevisiae homogenate . When the pH was up to 8.0, the SOD was adsorbed on the adsorbent. The adsorbed SOD was eluted by lowing pH and increasing ion strength. The recovery of SOD was 89% with purification factor 20 after elution with 0.5 mol / L NaCl citric buffer (pH4.5).