Zhang Pu, Fischer Amit, Ouyang Yu, Wang Jianbang, Sohn Yang Sung, Karmi Ola, Nechushtai Rachel, Willner Itamar
Institute of Chemistry, Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem Jerusalem 91904 Israel
Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University Chongqing 400715 People's Republic of China.
Chem Sci. 2022 Apr 29;13(25):7437-7448. doi: 10.1039/d2sc01542k. eCollection 2022 Jun 29.
Biomolecule-loaded nucleic acid-functionalized carboxymethyl cellulose hydrogel-stabilized microcapsules (diameter 2 μm) are introduced as cell-like containments. The microcapsules are loaded with two DNA tetrahedra, T and T, functionalized with guanosine-rich G-quadruplex subunits, and/or with native enzymes (glucose oxidase, GOx, and/or β-galactosidase, β-gal). In the presence of K-ions and hemin, the T/T tetrahedra constituents, loaded in the microcapsules, assemble into a hemin/G-quadruplex bridged tetrahedra dimer DNAzyme catalyzing the oxidation of Amplex Red to Resorufin by generating HO. In the presence of co-loaded GOx or GOx/β-gal, the GOx//T/T hemin/G-quadruplex cascade catalyzing the glucose-mediated oxidation of Amplex Red to Resorufin, and the three-biocatalysts cascade consisting of β-gal//GOx//hemin/G-quadruplex bridged T/T catalyzing the lactose-driven oxidation of Amplex Red to Resorufin proceed in the microcapsules. Enhanced biocatalytic transformations in the microcapsules, as compared to the performance of the reactions in a homogeneous phase, are observed, due to the proximity of the biocatalysts in a confined volume. As the synthetic methodology to prepare the microcapsules yields boundaries functionalized with complementary nucleic acid tethers, the dynamic association of different microcapsules, loaded selectively with biomolecular catalysts, proceeds. The dynamic dimerization of GOx-loaded microcapsules and hemin/G-quadruplex bridged T/T DNAzyme-loaded microcapsules yields effective intercommunicated microcapsules driving the GOx//hemin/G-quadruplex bridged T/T DNAzyme cascade. In addition, the dynamic dimerization of GOx-loaded microcapsules with β-gal//hemin/G-quadruplex bridged T/T-loaded microcapsules enables the bi-directional intercommunicated operation of the lactose-stimulated three catalysts β-gal//GOx//hemin/G-quadruplex bridged T/T DNAzyme cascade. The guided separation and formation of dynamic supramolecular dimer microcapsular containments, and the dictated switchable operation of intercommunicated biocatalytic cascades are demonstrated.
负载生物分子的核酸功能化羧甲基纤维素水凝胶稳定微胶囊（直径2μm）被用作类似细胞的容器。微胶囊中装载有两个用富含鸟苷的G-四链体亚基功能化的DNA四面体T和T，和/或天然酶（葡萄糖氧化酶，GOx，和/或β-半乳糖苷酶，β-gal）。在钾离子和血红素存在的情况下，微胶囊中装载的T/T四面体成分组装成血红素/G-四链体桥连的四面体二聚体DNAzyme，通过产生HO催化Amplex Red氧化为试卤灵。在共装载GOx或GOx/β-gal的情况下，GOx//T/T血红素/G-四链体级联催化葡萄糖介导的Amplex Red氧化为试卤灵，以及由β-gal//GOx//血红素/G-四链体桥连的T/T组成的三生物催化剂级联催化乳糖驱动的Amplex Red氧化为试卤灵在微胶囊中进行。与均相反应性能相比，由于生物催化剂在受限体积内的接近性，观察到微胶囊中的生物催化转化增强。由于制备微胶囊的合成方法产生了用互补核酸链功能化的边界，选择性装载生物分子催化剂的不同微胶囊的动态缔合得以进行。装载GOx的微胶囊和装载血红素/G-四链体桥连的T/T DNAzyme的微胶囊的动态二聚化产生了驱动GOx//血红素/G-四链体桥连的T/T DNAzyme级联的有效相互连通的微胶囊。此外，装载GOx的微胶囊与装载β-gal//血红素/G-四链体桥连的T/T的微胶囊的动态二聚化实现了乳糖刺激的三催化剂β-gal//GOx//血红素/G-四链体桥连的T/T DNAzyme级联的双向相互连通操作。展示了动态超分子二聚体微胶囊容器的引导分离和形成，以及相互连通的生物催化级联的可控切换操作。
