Nikolelis D P, Tzanelis M G, Krull U J
Department of Chemistry, University of Athens, Panepistimiopolis, Kouponia, Greece.
Biosens Bioelectron. 1994;9(3):179-88. doi: 10.1016/0956-5663(94)80119-3.
Bilayer lipid membranes (BLMs) can be used as generic transducers to monitor hydrolytic enzyme reactions occurring at the membrane surface. The representative enzymatic reactions presented herein were between membrane associated urease and penicillinase with urea and penicillin, respectively. Transient electrochemical signals from BLMs which contained enzyme were obtained by proper selection of the lipid composition of membranes. Negatively charged lipid membranes composed of egg phosphatidylcholine (PC) and 35% dipalmitoylphosphatidic acid were used for this purpose. The results were consistent with an electrostatic mechanism of perturbation of the surface structure of the BLMs, where changes of local hydronium ion activity associated with the enzymatic reaction altered the extent of ionization of the headgroups of the acidic constituent of the membranes, thereby providing a transient charging current which lasted for a period on the order of seconds. The delay time for observation of the transient was directly and reproducibly related to the concentration of the substrate which could be determined over a range of microM to mM levels. The results indicate that BLMs can be used as generic selective electrochemical transducers and as switchable biosensors to monitor rapid enzymatic reactions which alter pH.
双层脂质膜(BLMs)可用作通用传感器,以监测发生在膜表面的水解酶反应。本文介绍的代表性酶促反应分别是膜结合脲酶与尿素以及青霉素酶与青霉素之间的反应。通过适当选择膜的脂质组成,可以从含有酶的BLMs获得瞬态电化学信号。为此使用了由鸡蛋卵磷脂(PC)和35%二棕榈酰磷脂酸组成的带负电荷的脂质膜。结果与BLMs表面结构扰动的静电机制一致,其中与酶促反应相关的局部水合氢离子活性的变化改变了膜酸性成分头基团的电离程度,从而提供了持续约几秒的瞬态充电电流。观察到瞬态的延迟时间与底物浓度直接且可重复相关,底物浓度可在 microM 到 mM 水平范围内测定。结果表明,BLMs可用作通用选择性电化学传感器和可切换生物传感器,以监测改变pH值的快速酶促反应。