Purification of penicillin G acylase using immobilized metal affinity membranes.
The immobilized metal affinity membrane (IMAM) with modified regeneration cellulose was employed for purification of penicillin G acylase (PGA). For studying PGA adsorption capacity on the IMAM, factors such as chelator surface density, chelating metal, loading temperature, pH, NaCl concentration and elution solutions were investigated. The optimal loading conditions were found at 4 degrees C, 0.5 M NaCl, 32.04 micromol Cu(2+) per disk with 10 mM sodium phosphate buffer, pH 8.5, whereas elution conditions were: 1 M NH(4)Cl with 10 mM sodium phosphate buffer, pH 6.8. By applying these chromatographic conditions to the flow experiments in a cartridge, a 9.11-fold purification in specific activity with 90.25% recovery for PGA purification was obtained. Meanwhile, more than eight-times reusability of the membrane was achieved with the EDTA regeneration solutions.
Liu
Yung-Chuan
YC
Department of Chemical Engineering, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung, Taiwan 402. ycliu@dragon.nchu.edu.tw
ChangChien
Chih-Chiang
CC
Suen
Shing-Yi
SY
D000327:Adsorption; D002614:Chelating Agents / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation*; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D008670:Metals / Q000737:chemistry*; D010399:Penicillin Amidase / Q000302:isolation & purification*; D013696:Temperature
D002614:Chelating Agents; D008567:Membranes, Artificial; D008670:Metals; D010399:Penicillin Amidase