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用于在有机溶剂中分散和稳定有机磷水解酶的复杂凝聚核胶束。

Complex Coacervate Core Micelles for the Dispersion and Stabilization of Organophosphate Hydrolase in Organic Solvents.

机构信息

Department of Chemical Engineering, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts02139, United States.

出版信息

Langmuir. 2016 Dec 20;32(50):13367-13376. doi: 10.1021/acs.langmuir.6b02350. Epub 2016 Dec 7.

Abstract

Organophosphate (OP) nerve agents are a class of chemical warfare agents (CWAs) that exist as bulk stocks in current and past war zones. Thus, a technology that can perform on-site decontamination in a safe and timely fashion is desirable. Here, complex coacervate core micelles (C3Ms) were used to encapsulate organophosphate hydrolase (OPH) and chemostabilize it to maintain activity after exposure to organophosphate simulants ethanol and dimethyl methylphosphonate (DMMP). C3Ms were formed by two polymers-poly(acrylic acid) (PAA) and poly(oligo(ethylene glycol) methacrylate)-b-poly(4-vinyl N-methylpyridyl iodide), (POEGMA-b-qP4VP). Complexes of the coacervate micelles with the enzyme OPH were investigated by small angle neutron scattering (SANS), dynamic light scattering (DLS), and transmission electron microscopy (TEM), demonstrating the formation of micellar structures in solution. The activity of OPH against methyl paraoxon in these C3Ms under aqueous conditions was assayed after heat treatment for 3 days at 37 °C. The OPH in C3Ms retained 88 ± 7% of its initial activity, as compared to the 48 ± 3% activity retained by OPH alone, indicating that the C3Ms were able to stabilize the enzyme to heat treatment. C3Ms transferred into the two organic solvents formed larger structures than inverse micelles formed by the block copolymer alone. The addition of OPH to the C3Ms in organic solvents did not significantly change their structure. The activity of OPH (again, against methyl paraoxon) after 24 h of incubation at 4 °C was measured and compared to that of OPH in C3Ms. While OPH alone retained less than 5% of its activity after this incubation in both solvents, OPH in C3Ms retained 35 ± 3% of its activity in DMMP and 26 ± 1% of its activity in ethanol.

摘要

有机磷(OP)神经毒剂是一类化学战剂(CWA),目前和过去的战区都有大量库存。因此,需要一种能够安全、及时地进行现场去污的技术。在这里,我们使用复杂凝聚核胶束(C3M)来包封有机磷水解酶(OPH),并通过化学稳定化来维持其在暴露于有机磷模拟物乙醇和二甲基甲基膦酸酯(DMMP)后的活性。C3M 是由两种聚合物——聚丙烯酸(PAA)和聚(聚乙二醇甲基丙烯酸酯)-b-聚(4-乙烯基 N-甲基吡啶碘化物),(POEGMA-b-qP4VP)形成的。通过小角中子散射(SANS)、动态光散射(DLS)和透射电子显微镜(TEM)研究了凝聚胶束与酶 OPH 的复合物,证明了胶束结构在溶液中的形成。在 37°C 下热处理 3 天后,在这些 C3M 中用 OPH 对甲基对氧磷进行了活性测定。与单独的 OPH 相比,C3M 中的 OPH 保留了 88±7%的初始活性,表明 C3M 能够稳定酶以抵抗热处理。C3M 在两种有机溶剂中的转移形成的结构比单独的嵌段共聚物形成的反胶束更大。在有机溶剂中加入 OPH 不会显著改变它们的结构。在 4°C 下孵育 24 小时后,测量了 OPH(再次,针对甲基对氧磷)的活性,并与 C3M 中的 OPH 活性进行了比较。虽然单独的 OPH 在这两种溶剂中的孵育后保留了不到 5%的活性,但 C3M 中的 OPH 在 DMMP 中保留了 35±3%的活性,在乙醇中保留了 26±1%的活性。

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