Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrid, using a flexible linker to develop a more promising catalytic bioscavenger against a broad range of OPNA poisoning. GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNAs analogs (k_cat/K_m (10⁶ M^-1 min^-1): 59.8±0.06 (paraoxon), 5.2±0.02 (PNPDPP) and 47.0±6.0 (DFP)) and V-type OPNA analogs (k_cat/K_m (M^-1 min^-1): 504.3±48.5 (DSM) and 1324.0±47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.

中文翻译：

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工程重组 PON1-OPH 融合杂交体是一种潜在有效的催化生物清除剂，可对抗有机磷神经毒剂类似物。

有机磷神经毒剂 (OPNA)，包括 G 型和 V 型神经毒剂，如沙林、梭曼、塔崩和 VX，是具有极强神经毒性的有机磷化合物。由于化学解毒剂对 OPNA 中毒的保护作用低和不良副作用，因此正在开发能够水解 OPNA 的催化生物清除剂。然而，这些生物清除剂在实际应用中存在一定的局限性，包括催化活性低和特异性窄。在这项研究中，我们生成了一种融合杂交形式的工程重组人对氧磷酶 1 (rePON1) 和细菌有机磷水解酶 (OPH)，称为 GV 杂交，使用灵活的接头开发更有前途的催化生物清除剂，以对抗广泛的范围OPNA 中毒。对硝基苯基磷酸二苯酯 (PNPDPP)：2.3 ~ 33.0 倍和二异丙基氟磷酸盐 (DFP)：1.4 ~ 22.8 倍）和 V 型 OPNA 类似物（内吸子-S-甲基 (DSM)：1.9 ~ 34.6 倍和马拉硫磷: 1.1 ~ 4.2-fold above) 优于它们各自的酶形式。在 GV 杂交克隆中，GV7 克隆对两种 G 型 OPNAs 类似物的催化活性都有显着改善（k _cat / K _m（10 ⁶ M ^-1 min ^-1）：59.8±0.06（对氧磷），5.2± 0.02 (PNPDPP) 和 47.0±6.0 (DFP)) 和 V 型 OPNA 类似物 ( k _cat / K _m (M ^-1 min ^-1): 504.3±48.5 (DSM) 和 1324.0±47.5 (马拉硫磷))。总之，我们开发了 GV 杂交形式的 rePON1 和细菌 OPH 突变体作为有效且合适的催化生物清除剂来水解广泛的 OPNA 类似物。