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首页> 外文学位 >Bifunctional fusion proteins for the detoxification and monitoring of organophosphate pesticides: Surface expression and environmentally triggered purification/immobilization.
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Bifunctional fusion proteins for the detoxification and monitoring of organophosphate pesticides: Surface expression and environmentally triggered purification/immobilization.

机译：用于解毒和监测有机磷酸酯农药的双功能融合蛋白：表面表达和环境触发的纯化/固定化。

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摘要

Organophosphorus nerve agents are highly toxic compounds that are widely used as pesticides and chemical warfare agents. Novel bifunctional fusion proteins were created to address the shortcomings of current bioremediation and detection strategies. In one example, whole cell biocatalysts were generated by targeting a detoxification enzyme, organophosphate hydrolase (OPH), onto the cell surface in order to alleviate the potential diffusional limitations created by the cell membrane. Using an ice-nucleation protein (INP) anchor, OPH was successfully localized onto the cell surface of Escherichia coli, Pseudomonas sp. and Moraxella sp., resulting in whole cell biocatalysts that rapidly hydrolyzed organophosphate nerve agents. Utilizing Moraxella sp. and it's natural p-nitrophenol degradative pathway, simultaneous degradation of the pesticide as well as it's hydrolysis product was achieved for complete mineralization of selected pesticides.; A bifunctional fusion protein capable of purifying OPH without the use of conventional chromatography was also described. OPH was fused to a thermally responsive biopolymer, elastin-like protein (ELP). The ELP fusion enabled rapid purification of the OPH fusions with simple environmental triggers. The fusion protein retained nearly identical kinetics as the wild-type OPH. In addition the ELP domain served as an immobilization tag onto hydrophobic supports. By creating a spatially resolved temperature profile, a simple and reversible patterning technique can be achieved on an inexpensive substrate by thermally addressing a patterned hydrophobic surface template.
机译：有机磷神经毒剂是剧毒化合物，被广泛用作农药和化学战剂。创建了新型的双功能融合蛋白，以解决当前生物修复和检测策略的缺点。在一个实例中，通过将解毒酶有机磷酸水解酶（OPH）靶向到细胞表面上来产生全细胞生物催化剂，以减轻由细胞膜产生的潜在扩散限制。使用冰核蛋白（INP）锚，将OPH成功定位到大肠杆菌Pseudomonas sp的细胞表面上。和 Moraxella sp。，导致全细胞生物催化剂迅速水解有机磷酸盐神经毒剂。利用 Moraxella sp。它是天然的对硝基苯酚降解途径，可同时降解农药及其水解产物，从而使选定农药完全矿化。还描述了一种无需使用常规色谱即可纯化OPH的双功能融合蛋白。 OPH与热响应性生物聚合物弹性蛋白样蛋白（ELP）融合。 ELP融合能够通过简单的环境触发因素快速纯化OPH融合。融合蛋白保留了与野生型OPH几乎相同的动力学。另外，ELP结构域用作疏水性支持物上的固定标签。通过创建空间分辨的温度曲线，可以通过对寻址的疏水表面模板进行热寻址，在廉价的基板上实现简单且可逆的构图技术。

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  • 作者

    Shimazu, Mark Koichi.;

  • 作者单位

    University of California, Riverside.;

  • 授予单位 University of California, Riverside.;
  • 学科 Engineering Environmental.; Chemistry Agricultural.
  • 学位 Ph.D.
  • 年度 2003
  • 页码 107 p.
  • 总页数 107
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 环境污染及其防治;农业化学;
  • 关键词

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