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Combinatorial Strategy for Studying Biochemical Pathways in Double Emulsion Templated Cell‐Sized Compartments
Author(s) -
Santos Elena C.,
Belluati Andrea,
Necula Danut,
Scherrer Dominik,
Meyer Claire E.,
Wehr Riccardo P.,
Lörtscher Emanuel,
Palivan Cornelia G.,
Meier Wolfgang
Publication year - 2020
Publication title -
advanced materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 10.707
H-Index - 527
eISSN - 1521-4095
pISSN - 0935-9648
DOI - 10.1002/adma.202004804
Subject(s) - interconnectivity , microfluidics , membrane , cascade , materials science , nanotechnology , enzyme , artificial cell , biophysics , chemistry , biological system , biology , biochemistry , computer science , chromatography , artificial intelligence
Cells rely upon producing enzymes at precise rates and stoichiometry for maximizing functionalities. The reasons for this optimal control are unknown, primarily because of the interconnectivity of the enzymatic cascade effects within multi‐step pathways. Here, an elegant strategy for studying such behavior, by controlling segregation/combination of enzymes/metabolites in synthetic cell‐sized compartments, while preserving vital cellular elements is presented. Therefore, compartments shaped into polymer GUVs are developed, producing via high‐precision double‐emulsion microfluidics that enable: i) tight control over the absolute and relative enzymatic contents inside the GUVs, reaching nearly 100% encapsulation and co‐encapsulation efficiencies, and ii) functional reconstitution of biopores and membrane proteins in the GUVs polymeric membrane, thus supporting in situ reactions. GUVs equipped with biopores/membrane proteins and loaded with one or more enzymes are arranged in a variety of combinations that allow the study of a three‐step cascade in multiple topologies. Due to the spatiotemporal control provided, optimum conditions for decreasing the accumulation of inhibitors are unveiled, and benefited from reactive intermediates to maximize the overall cascade efficiency in compartments. The non‐system‐specific feature of the novel strategy makes this system an ideal candidate for the development of new synthetic routes as well as for screening natural and more complex pathways.