Immobilization of CYP1A2 and CYP Reductase onto Self-assembled Phospholipid Layer and Evaluation of their Activity

Abstract

We had immobilized cytochrome P450 (CYP) 1A2 and CYP reductase (CPR) onto a self-assembled phospholipid layer containing stearic acid (LDPE-StA-PC-SA) to confirm the functional interaction between CYP 1A2 and CPR. The formation of resorufin from 7-ethoxy resorufin was observed by use of the film immobilizing CYP 1A2 and CPR onto LDPE-StA-PC-SA. It was clarified that the fluidity of LDPE-StA-PC-SA was very important. The actual activity of CYP 1A2 did not depend on the density of CYP 1A2 on LDPE-StA-PC-SA, although its specific activity was affected on its density. The activity of immobilized CYP 1A2 depended on a temperature. It was assumed that the optimal temperature was about 45 ℃.