Construction of coacervate-in-coacervate multi-compartment protocells for spatial organization of enzymatic reactions.
Construction of coacervate-in-coacervate multi-compartment protocells for spatial organization of enzymatic reactions.
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用于酶反应空间组织的凝聚层中凝聚层多室原细胞的构建
DOI：
10.1039/d0sc03849k
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发表时间：
2020-08-07
期刊：
影响因子：
8.4
通讯作者：
Liu J
中科院分区：
文献类型：
--
作者：
Chen Y;Yuan M;Zhang Y;Liu S;Yang X;Wang K;Liu J
通过液-液相分离形成的凝聚层微滴已经被广泛探索作为用于构建人工细胞或细胞器的隔室模型。在这项研究中，凝聚层中凝聚层多室原始细胞的构建使用四种聚电解质，其中羧甲基葡聚糖和二乙基氨基乙基葡聚糖沉积在所制备的聚二烯丙基二甲基铵/脱氧核糖核酸凝聚层微滴的表面通过层层组装。所得到的多室原始细胞由两个不混溶的凝聚相组成，具有不同的物理和化学性质。分子传输实验表明，小分子可以在两个凝聚相之间扩散，大分子酶可以保留。此外，在多室原始细胞中进行葡萄糖氧化酶/辣根过氧化物酶-过氧化氢酶的竞争性级联酶促反应。不同的酶组织和H2 O2的产生导致多巴胺的不同聚合。不同的酶在不混溶的凝聚相，凝聚相之间的不同的反应通量，和酶级联网络的空间组织导致可区分的信号产生和产品输出。这种多隔室结构的发展可能为多酶级联的空间组织铺平道路，并为含细胞器的人工细胞的设计提供新的思路。凝聚层中凝聚层的微结构被构建为多隔室原始细胞模型，其中多酶级联在空间上被组织用于竞争性酶促反应。
Coacervate microdroplets, formed via liquid–liquid phase separation, have been extensively explored as a compartment model for the construction of artificial cells or organelles. In this study, coacervate-in-coacervate multi-compartment protocells were constructed using four polyelectrolytes, in which carboxymethyl-dextran and diethylaminoethyl-dextran were deposited on the surface of as-prepared polydiallyldimethyl ammonium/deoxyribonucleic acid coacervate microdroplets through layer-by-layer assembly. The resulting multi-compartment protocells were composed from two immiscible coacervate phases with distinct physical and chemical properties. Molecule transport experiments indicated that small molecules could diffuse between two coacervate phases and that macromolecular enzymes could be retained. Furthermore, a competitive cascade enzymatic reaction of glucose oxidase/horseradish peroxidase–catalase was performed in the multi-compartment protocells. The different enzyme organization and productions of H2O2 led to a distinct polymerization of dopamine. The spatial organization of different enzymes in immiscible coacervate phases, the distinct reaction fluxes between coacervate phases, and the enzymatic cascade network led to distinguishable signal generation and product outputs. The development of this multi-compartment structure could pave the way toward the spatial organization of multi-enzyme cascades and provide new ideas for the design of organelle-containing artificial cells. A coacervate-in-coacervate micro-architecture is constructed as a multi-compartment protocell model, in which a multi-enzyme cascade is spatially organized for competitive enzymatic reactions.
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影响因子：
15
作者：
Fu, Jingcheng;Schlenoff, Joseph B.
通讯作者：
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作者：
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