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.
Artificial organelles created from a bottom up approach are a new type of engineered materials, which are not designed to be living but, instead, to mimic some specific functions inside cells. By doing so, artificial organelles are expected to become a powerful tool in biomedicine. They can act as nanoreactors to convert a prodrug into a drug inside the cells or as carriers encapsulating therapeutic enzymes to replace malfunctioning organelles in pathological conditions. For the design of artificial organelles, several requirements need to be fulfilled: a compartmentalized structure that can encapsulate the synthetic machinery to perform an enzymatic function, as well as a means to allow for communication between the interior of the artificial organelle and the external environment, so that substrates and products can diffuse in and out the carrier allowing for continuous enzymatic reactions. The most recent and exciting advances in architectures that fulfill the aforementioned requirements are featured in this review. Artificial organelles are classified depending on their constituting materials, being lipid and polymer-based systems the most prominent ones. Finally, special emphasis will be put on the intracellular response of these newly emerging systems.
Phenylketonuria (PKU) is a genetic enzyme defect affecting 1:10 000–20 000 newborn children every year. The amino acid phenylalanine (Phe) is not depleted but accumulates in tissues of several organs, which leads to severe medical conditions. A promising concept to restore the metabolism of the affected patients will be to orally administer the defective enzyme which will remove Phe in the intestine. Herein, capsosomes, a multicompartment carrier consisting of thousands of liposomes embedded within a polymeric carrier, are employed as encapsulation platform for this purpose. It is shown that the enzyme phenylalanine ammonia lyase can be entrapped within the liposomal compartments with preserved activity, demonstrated by the conversion of Phe into trans‐cinnamic acid (t‐ca). With the aim to mimic the dynamic environment in the intestine, the Phe conversion is performed in a microfluidic set up in the presence of human intestinal epithelial cells with applied intestinal flow and peristaltic motions. It is also shown that the microreactors are neither internalized by the cells nor exhibit inherent cytotoxicity while concurrently converting Phe into t‐ca. Taken together, the first active extracellular multicompartment microreactor is reported using the relevant enzymes and settings toward the treatment of the medical condition PKU.
A fungal endoxylanase belonging to the glycoside hydrolase gene family 11 (GH11) was obtained from the ascomycete Talaromyces amestolkiae. The enzyme was purified, characterized and used to produce a mixture of xylooligosaccharides (XOS) from birchwood xylan. A notable yield of neutral XOS was obtained (28.8%) upon enzyme treatment and the mixture contained a negligible amount of xylose, having xylobiose, xylotriose and xylotetraose as its main components. The prebiotic potential of this mixture was demonstrated upon analyzing the variations in microorganisms' composition and organic acids profile in breast-fed child faeces fermentations. The strong production of acetic and lactic acid, the decrease of potentially pathogenic bacteria and the increase of bifidobacteria, and possible beneficial commensals, confirmed the prebiotic value of these xylooligosaccharides.
Elevated ROS levels are related to the initiation and progression of many severe diseases. Herein, we present for the first time a microreactor conducting non-enzymatic and enzymatic activity for the depletion of ROS.
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