Various immobilized metal ions affinity membranes (IMAMs) were prepared from the regenerated cellulosemembrane (RC membrane) and chelated with various metal ions such as Co2+, Ni2+, Cu2+ and Zn2+.The D-hydantoin-hydrolyzing enzyme (DHTase) harboring a poly-His tagged residue was used as a modelprotein to be immobilized on the prepared IMAMs through the direct metal–protein interaction forces.The adsorption isotherm and the kinetic parameters Vmax, Km,app of DHTase on IMAMs were studied. Thecobalt ions chelated IMAM (Co-IMAM) was found to yield the highest specific activity of DHTase. Underthe immobilization condition, the cobalt ion chelated amount was 161.4 ± 4.7 mol/disk with a DHTaseactivity of 4.1 ± 0.1 U/disk. As compared to the free DHTase, the immobilized DHTase membrane couldachieve a broader pH tolerance and higher thermal stability. In addition, 98% of the residual activitycould be retained for 7-times repeated use. Only little activity loss was observed within 36-day storageat 4 ◦C. This is the first report concerning about using cobalt ion as the effective chelated metal ion forsimultaneous purification and immobilization operatio
To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.