Masson P, Josse D, Lockridge O, Viguié N, Taupin C, Buhler C
Centre de Recherches du Service de Santé des Armées, Department of Toxicology, La Tronche, France.
J Physiol Paris. 1998 Oct-Dec;92(5-6):357-62. doi: 10.1016/S0928-4257(99)80005-9.
Enzymes hydrolyzing organophosphates could be used as catalytic scavengers for treatment of organophosphate poisoning and for decontamination. Two organophosphorus hydrolases (OPH) were selected: the Flavobacterium sp/Pseudomonas diminuta phosphotriesterase (PTE) and human paraoxonase (HuPON). Genes encoding these enzymes were cloned and functional recombinant enzymes expressed. PTE was expressed in E. coli. Natural HuPON was purified from human plasma; recombinant HuPON was expressed in human embryonic kidney 293 T cells. Although HuPON displays interesting catalytic properties, a site-directed mutagenesis program was undertaken to improve its catalytic efficiency. PTE has high efficiency in hydrolysis of organophosphates, including nerve agents. PTE injected in rat has a half-life of 100 min. However, to overcome pharmacokinetic problems of injected OPH and/or immunological incompatibility, the model enzyme (recombinant PTE) was immobilized onto a hollow-fiber reactor. This reactor designed for extracorporeal blood circulation is under experimentation for post-exposure detoxification.
水解有机磷酸酯的酶可用作催化清除剂，用于治疗有机磷酸酯中毒和去污。选择了两种有机磷水解酶（OPH）：黄杆菌属/微小假单胞菌磷酸三酯酶（PTE）和人对氧磷酶（HuPON）。克隆了编码这些酶的基因并表达了功能性重组酶。PTE在大肠杆菌中表达。天然HuPON从人血浆中纯化；重组HuPON在人胚肾293 T细胞中表达。尽管HuPON具有有趣的催化特性，但仍开展了定点诱变计划以提高其催化效率。PTE在水解包括神经毒剂在内的有机磷酸酯方面具有高效性。注射到大鼠体内的PTE半衰期为100分钟。然而，为了克服注射用OPH的药代动力学问题和/或免疫不相容性，将模型酶（重组PTE）固定在中空纤维反应器上。这种设计用于体外血液循环的反应器正在进行暴露后解毒实验。
