Quantitative evaluation of luminescence intensity from enzymatic luminescence reaction of coelenterazine and analogues

Yuma Ikeda, Masanobu Tanaka, Ryo Nishihara, Yuki Hiruta, Daniel Citterio, Koji Suzuki, Kazuki Niwa

研究成果: Article査読

7 被引用数 (Scopus)

抄録

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.

本文言語English
論文番号112459
ジャーナルJournal of Photochemistry and Photobiology A: Chemistry
394
DOI
出版ステータスPublished - 2020 5月 1

ASJC Scopus subject areas

  • 化学一般
  • 化学工学一般
  • 物理学および天文学一般

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