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Confined Multiple Enzymatic (Cascade) Reactions within Poly(dopamine)-based Capsosomes

机译:基于聚(多巴胺)的脂质体中的受限多个酶促反应(级联)

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The design of compartmentalized carriers as artificial cells is envisioned to be an efficient tool with potential applications in the biomedical field. The advent of this area has witnessed the assembly of functional, bioinspired systems attempting to tackle challenges in cell mimicry by encapsulating multiple compartments and performing controlled encapsulated enzymatic catalysis. Although capsosomes, which consist of liposomes embedded within a polymeric carrier capsule, are among the most advanced systems, they are still amazingly simple in their functionality and cumbersome in their assembly. We report on capsosomes by embedding liposomes within a poly(dopamine) (PDA) carrier shell created in a solution-based single-step procedure. We demonstrate for the first time the potential of PDA-based capsosomes to act as artificial cell mimics by performing a two-enzyme coupled reaction in parallel with a single-enzyme conversion by encapsulating three different enzymes into separated liposomal compartments. In the former case, the enzyme uricase converts uric acid into hydrogen peroxide, CO2 and allantoin, followed by the reaction of hydrogen peroxide with the reagent Amplex Ultra Red in the presence of the enzyme horseradish peroxidase to generate the fluorescent product resorufin. The parallel enzymatic catalysis employs the enzyme ascorbate oxidase to convert ascorbic acid into 2-L-dehydroascorbic acid.
机译:设想将分隔的载体设计为人工细胞是一种有效的工具,在生物医学领域具有潜在的应用。该领域的出现见证了功能性,受生物启发的系统的组装,这些系统试图通过封装多个隔室并执行受控的封装酶催化来应对细胞模仿中的挑战。尽管由包裹在聚合物载体胶囊中的脂质体组成的脂质体是最先进的系统,但它们的功能仍然非常简单,而且组装麻烦。我们通过在基于溶液的单步程序中创建的聚(多巴胺)(PDA)载体壳中嵌入脂质体来报告脂质体。我们首次通过将两种不同的酶封装到分开的脂质体区室中,通过与单酶转化同时进行两种酶偶联反应,证明了基于PDA的脂质体作为人工细胞模拟物的潜力。在前一种情况下,尿酸酶将尿酸转化为过氧化氢,CO2和尿囊素,然后在辣根过氧化物酶的存在下使过氧化氢与试剂Amplex Ultra Red反应生成荧光产物试卤灵。平行酶催化利用抗坏血酸氧化酶将抗坏血酸转化为2-L-脱氢抗坏血酸。

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