Immobilized IDA-type Cu2+-chelating membrane affinity chromatography for purification of bovine liver catalase.
Cu2+-chelating membrane medium based on iminodiacetate-substituted modified short cotton cellulose was examined to purify bovine liver catalase (BLC) by affinity chromatography. The buffer pH notably affect BLC binding to the Cu2+-chelating membrane adsorbent. The results showed that under the chromatographic conditions chosen, 67.7% recovery of BLC attained with an overall 4.7-fold increase in specific activity in a single step. After purification of BLC, the chelating membrane adsorbent can easily be regenerated by imidazole or EDTA buffer. Different results were obtained by the two regeneration methods: the later is more effective than the former in reviving the purification characters of the chelating membrane adsorbent.
Yang
L
L
National Chromatographic R. & A. Center of China, Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian, 116011.
Jia
L
L
Zou
H
H
Kong
L
L
Zhang
Y
Y
D000818:Animals; D002374:Catalase / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D003300:Copper / Q000737:chemistry; D008099:Liver / Q000737:chemistry / Q000201:enzymology*; D008567:Membranes, Artificial; D011485:Protein Binding
D002614:Chelating Agents; D008567:Membranes, Artificial; D003300:Copper; D002374:Catalase