Supercharged enzyme-polymer surfactant bioblocks for the preparation of organophosphate decontaminating materials

用于制备有机磷去污材料的增压酶聚合物表面活性剂生物嵌段

• 批准号: EP/N026586/1
• 负责人: Adam Perriman
• 金额: $ 45.53万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FN026586%2F1
• 关键词: Supercharged; enzyme; polymer; surfactant; bioblocks

Since their widespread application as pesticides in rural areas of developing countries, it is estimated that approximately 3 million people worldwide are poisoned by organophosphates (OPs) every year. OPs have also been used in chemical warfare agent formulations, in incidents including the Ghouta Sarin Attack in Syria, 2013, and the Japanese subway attacks in 1994 and 1995. According to a French intelligence assessment published in September 2013, stockpiles in Syria alone include several hundreds of tonnes of sarin and several tens of tonnes of VX. Accordingly, this research proposal describes the application of synthetic biology for the rational design of versatile supercharged enzyme-polymer surfactant building blocks (bioblocks) for the preparation of organophosphate decontaminating materials that span all three phases of matter. Here, the surfaces of synthetic supercharged variants of the organophosphate-degrading enzyme organophosphate hydrolase (OPH) will be radically re-engineered to produce adhesive enzyme-polymer melts, stable bioaerosols, and hierarchically assembled solid membranes.The versatility in this new methodology is highly dependent on the ability to manipulate protein-protein interactions through the construction of an electrostatically-assembled polymer surfactant corona at the surface of a supercharged enzyme, which is based on the synthetic methodology recently pioneered by AWP. The approach involves the reengineering of a protein surface in two key steps: (i) amplification of the positive charge density on the protein surface, followed by (ii) electrostatic coupling of anionic polymer surfactant chains to the cationic sites on the protein surface. Significantly, the resulting surface-bound corona of polymer surfactant molecules increases the range of the attractive intermolecular protein-protein interactions, which in turn allows the particle motions required for melt formation under anhydrous conditions. Alternatively, the hydrophilic-lipophilic balance (HLB) of the corona can be tuned to either provide organic solvent compatibility for aerosol generation or to promote surfactant mediated self-assembly to produce nanoporous solids. Accordingly, the global aim of this research proposal is the rational design and synthesis of the organophosphate-degrading enzyme-polymer surfactant bioblocks that can be used for the preparation of these three classes of organophosphate decontaminating materials. The research program will be implemented sequentially across four primary research objectives: - In silico inspired design, expression and purification of a supercharged organophosphate hydrolase (scOPH) library - The synthesis of high-density scOPH-polymer melts - Active bioaerosol generation using organic solvent compatibility - Surfactant-mediated assembly of scOPH to give porous solids with recyclable catalytic activitiesThe research programme describes a scientific approach that combines in-house techniques for synthetic biology, biophysics and materials science, as well as techniques available at large-scale facilities. As there is a strong application focus in the programme, the new methodology describes the development of recombinant supercharged enzymes, which will be optimised for maximum catalytic performance. In conclusion, the development of a library of OP-degrading enzyme-polymer surfactant materials that can operate in all three phases represents a near-future platform technology that could be readily exploited for a multitude of new defence applications, including disbondable coating for military hardware or personnel, bioaerosol-based countermeasures for OP contaminated confined airspaces or for inhalation treatments, and high efficiency enzyme-based reactors for OP degradation/disposal.

由于有机磷农药在发展中国家农村地区的广泛应用，据估计，全世界每年约有300万人因有机磷农药中毒。OP还被用于化学战剂配方，包括2013年叙利亚Ghouta Sarin袭击事件和1994年和1995年日本地铁袭击事件。根据2013年9月发布的法国情报评估，仅叙利亚的库存就包括数百吨沙林和数十吨VX。因此，本研究建议描述了合成生物学的应用，用于合理设计通用的增压酶-聚合物表面活性剂构建块（生物块），用于制备跨越所有三个物质相的有机磷酸盐去污材料。在这里，有机磷降解酶有机磷水解酶（OPH）的合成增压变体的表面将被彻底重新设计，以产生粘性酶-聚合物熔体，稳定的生物气溶胶，和分级组装的固体膜。这种新方法的多功能性高度依赖于通过构建静电-组装的聚合物表面活性剂电晕在表面的超荷电酶，这是基于合成方法最近开创的AWP。该方法涉及两个关键步骤中的蛋白质表面的再工程：（i）蛋白质表面上的正电荷密度的放大，随后（ii）阴离子聚合物表面活性剂链与蛋白质表面上的阳离子位点的静电偶联。值得注意的是，所得到的聚合物表面活性剂分子的表面结合的冠增加了有吸引力的分子间蛋白质-蛋白质相互作用的范围，这反过来又允许在无水条件下熔融形成所需的颗粒运动。或者，可以调节电晕的亲水亲油平衡（HLB）以提供用于气溶胶生成的有机溶剂相容性或促进表面活性剂介导的自组装以产生纳米多孔固体。因此，本研究计划的总体目标是合理设计和合成可用于制备这三类有机磷去污材料的有机磷降解酶-聚合物表面活性剂生物块。该研究计划将在四个主要研究目标中依次实施：- 在silico灵感的设计，超荷电有机磷酸水解酶（scOPH）文库的表达和纯化-高密度scOPH-聚合物熔体的合成-使用有机溶剂相容性的活性生物气溶胶产生-表面活性剂-介导组装scOPH以提供具有可回收催化活性的多孔固体该研究计划描述了一种科学方法，该方法结合了合成生物学的内部技术，生物物理学和材料科学，以及在大型设施中可用的技术。由于该计划有很强的应用重点，新方法描述了重组增压酶的开发，这些酶将被优化以获得最大的催化性能。总之，可以在所有三个阶段发挥作用的OP降解酶-聚合物表面活性剂材料库的开发代表了一种不久的将来的平台技术，可以很容易地用于多种新的国防应用，包括用于军事硬件或人员的可剥离涂层，针对OP污染的密闭空气空间或吸入治疗的基于生物气溶胶的对策，以及用于OP降解/处置的高效酶基反应器。

项目成果

Measure of porosity in flax fibres reinforced polylactic acid biocomposites

• DOI: 10.1016/j.compositesa.2020.106183
• 发表时间: 2021-02-01
• 期刊: COMPOSITES PART A-APPLIED SCIENCE AND MANUFACTURING
• 影响因子: 8.7
• 作者: de Kergariou, Charles;Le Duigou, Antoine;Scarpa, Fabrizio
• 通讯作者: Scarpa, Fabrizio

Hydrogel-Immobilized Supercharged Proteins

• DOI: 10.1002/adbi.201700240
• 发表时间: 2018-07-01
• 期刊: ADVANCED BIOSYSTEMS
• 影响因子: 4.1
• 作者: Campbell, Eleanor C.;Grant, Jacob;Jackson, Colin J.
• 通讯作者: Jackson, Colin J.