Optimizing the Chemiluminescence of a Light-Producing Deoxyribozyme

0556678 - ÚOCHB 2023 RIV DE eng J - Journal Article
Jakubec, Martin - Pšenáková, Karolína - Švehlová, Kateřina - Curtis, Edward A.
Optimizing the Chemiluminescence of a Light-Producing Deoxyribozyme.
Chembiochem. Roč. 23, č. 10 (2022), č. článku e202200026. ISSN 1439-4227. E-ISSN 1439-7633
R&D Projects: GA ČR(CZ) GA19-20989S
Institutional support: RVO:61388963
Keywords : chemiluminescence * deoxyribozyme * in vitro selection * sensors * supernova
OECD category: Biochemistry and molecular biology
Impact factor: 3.2, year: 2022
Method of publishing: Limited access
https://doi.org/10.1002/cbic.202200026

Supernova is a chemiluminescent deoxyribozyme recently discovered in our group. It transfers the phosphate group from the 1,2-dioxetane substrate CDP-Star to its 5 ' hydroxyl group, which triggers a decomposition reaction and the production of light. Here we investigated the effects of reaction conditions on the ability of Supernova to generate a chemiluminescent signal (using a plate reader assay) and to phosphorylate itself (using a ligation assay). Our experiments indicate that multiple zinc ions are required for catalytic function, suggesting links between Supernova and protein enzymes that catalyze similar reactions. They also show how factors such as pH, potassium concentration, CDP-Star concentration, and DNA concentration affect the reaction. By combining information from different experiments, the rate enhancement of light production was increased by more than 1000-fold. These results should be useful for applications in which Supernova is used as a sensor.
Permanent Link: http://hdl.handle.net/11104/0330794