Article: Electroanalysis at a Single Giant Vesicle Generating Enzymatically a Reactive Oxygen Species
Analytical chemistry. 2021 Sept. 21, v. 93, no. 39
2021
Abstract: In the framework of artificial or synthetic cell development, giant liposomes are common basic structures. Their enclosed membrane allows encapsulating proteins, DNA, reactants, etc., while its phospholipid nature allows some exchanges with the ... ...
| Abstract | In the framework of artificial or synthetic cell development, giant liposomes are common basic structures. Their enclosed membrane allows encapsulating proteins, DNA, reactants, etc., while its phospholipid nature allows some exchanges with the surrounding medium. Biochemical reactions induced inside giant liposomes or vesicles are often monitored or imaged by fluorescence microscopy techniques. Here, we show that electrochemistry performed with ultramicroelectrodes is perfectly suitable to monitor an enzymatic reaction occurring in a single giant unilamellar vesicle. Glucose oxidase (GOx) was microinjected inside individual vesicles containing 1 mM glucose. H₂O₂ was thus generated in the vesicle and progressively diffused across the membrane toward the surrounding environment. An ultramicroelectrode sensitive to H₂O₂ (black platinum-modified carbon surface) was placed next to the membrane and provided a direct detection of the hydrogen peroxide flux generated by the enzyme activity. Electrochemistry offered a highly sensitive (in situ detection), selective (potential applied at the electrode), time-resolved analysis (chronoamperometry) of the GOx activity over an hour duration, without modifying the internal giant unilamellar vesicles (GUV) medium. These results demonstrate that electroanalysis with microsensors is well adapted and complementary to fluorescence microscopy to sense enzymatic activities, for instance, generating reactive oxygen species, at single vesicles further used to develop artificial cells. |
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| Keywords | DNA ; analytical chemistry ; artificial cells ; carbon ; electrochemistry ; electrodes ; enzymatic reactions ; enzyme activity ; fluorescence microscopy ; glucose ; glucose oxidase ; hydrogen peroxide ; phospholipids |
| Language | English |
| Dates of publication | 2021-0921 |
| Size | p. 13143-13151. |
| Publishing place | American Chemical Society |
| Document type | Article |
| ZDB-ID | 1508-8 |
| ISSN | 1520-6882 ; 0003-2700 |
| ISSN (online) | 1520-6882 |
| ISSN | 0003-2700 |
| DOI | 10.1021/acs.analchem.1c01208 |
| Database | NAL-Catalogue (AGRICOLA) |
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