2020年5月1日
Quantitative evaluation of luminescence intensity from enzymatic luminescence reaction of coelenterazine and analogues
Journal of Photochemistry and Photobiology A: Chemistry
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- 巻
- 394
- 号
- 開始ページ
- 112459
- 終了ページ
- 112459
- 記述言語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1016/j.jphotochem.2020.112459
- 出版者・発行元
- Journal of Photochemistry and Photobiology A: Chemistry
© 2020 Elsevier B.V. Recent advances in bioluminescence technology rely on the discovery of luciferin-luciferase pairs having excellent luminescence activity. Here, we focus on coelenterazine (CTZ) and its analogues and quantitatively investigate reaction factors, including bioluminescence reaction quantum yield (φBL) and enzyme reaction kinetics (turnover rate, kcat), determining the luminescence intensity to design brighter reaction systems. Initially, it was confirmed that the CTZ was kept stable during the storage and the φBL value measuring experiment in terms of comparative study using benzyl-protected CTZ. Using a luminometer whose absolute responsivity is calibrated for each luminescence reaction spectrum, we absolutely measured φBL and other reaction factors of CTZ and 3 types of blue-shifted CTZ analogues, CTZ 400a, 6-pi-H-CTZ and 6-pi-Ph-CTZ. The results suggested that there was no significant difference in reaction kinetics, but a clear difference in φBL (up to 22-fold), which was presumably due to an improvement in fluorescence quantum yield (φFL) of the corresponding coelenteramide (CTMD), oxidized form of CTZ. Our evaluation suggests that the design of CTZ analogues considering φFL is important for developing a bright luminescent reaction system.
- リンク情報
- ID情報
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- DOI : 10.1016/j.jphotochem.2020.112459
- ISSN : 1010-6030