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A Minimized Chemoenzymatic Cascade for Bacterial Luciferase in Bioreporter Applications
Author(s) -
Phonbuppha Jittima,
Tinikul Ruchanok,
Wongnate Thanyaporn,
Intasian Pattarawan,
Hollmann Frank,
Paul Caroline E.,
Chaiyen Pimchai
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000100
Subject(s) - flavin group , bioluminescence , luciferase , cascade reaction , reductase , biosensor , chemistry , combinatorial chemistry , biochemistry , luciferin , cofactor , flavin mononucleotide , redox , biocatalysis , enzyme , catalysis , gene , transfection , organic chemistry , reaction mechanism
Bacterial luciferase (Lux) catalyzes a bioluminescence reaction by using long‐chain aldehyde, reduced flavin and molecular oxygen as substrates. The reaction can be applied in reporter gene systems for biomolecular detection in both prokaryotic and eukaryotic organisms. Because reduced flavin is unstable under aerobic conditions, another enzyme, flavin reductase, is needed to supply reduced flavin to the Lux‐catalyzed reaction. To create a minimized cascade for Lux that would have greater ease of use, a chemoenzymatic reaction with a biomimetic nicotinamide (BNAH) was used in place of the flavin reductase reaction in the Lux system. The results showed that the minimized cascade reaction can be applied to monitor bioluminescence of the Lux reporter in eukaryotic cells effectively, and that it can achieve higher efficiencies than the system with flavin reductase. This development is useful for future applications as high‐throughput detection tools for drug screening applications.