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- Title
Bioluminescence of (R)-Cypridina Luciferin with Cypridina Luciferase.
- Authors
Kanie, Shusei; Wu, Chun; Kihira, Kiyohito; Yasuno, Rie; Mitani, Yasuo; Ohmiya, Yoshihiro
- Abstract
Cypridina luciferin (CypL) is a marine natural product that functions as the luminous substrate for the enzyme Cypridina luciferase (CypLase). CypL has two enantiomers, (R)- and (S)-CypL, due to its one chiral center at the sec-butyl moiety. Previous studies reported that (S)-CypL or racemic CypL with CypLase produced light, but the luminescence of (R)-CypL with CypLase has not been investigated. Here, we examined the luminescence of (R)-CypL, which had undergone chiral separation from the enantiomeric mixture, with a recombinant CypLase. Our luminescence measurements demonstrated that (R)-CypL with CypLase produced light, indicating that (R)-CypL must be considered as the luminous substrate for CypLase, as in the case of (S)-CypL, rather than a competitive inhibitor for CypLase. Additionally, we found that the maximum luminescence intensity from the reaction of (R)-CypL with CypLase was approximately 10 fold lower than that of (S)-CypL with CypLase, but our kinetic analysis of CypLase showed that the Km value of CypLase for (R)-CypL was approximately 3 fold lower than that for (S)-CypL. Furthermore, the chiral high-performance liquid chromatography (HPLC) analysis of the reaction mixture of racemic CypL with CypLase showed that (R)-CypL was consumed more slowly than (S)-CypL. These results indicate that the turnover rate of CypLase for (R)-CypL was lower than that for (S)-CypL, which caused the less efficient luminescence of (R)-CypL with CypLase.
- Subjects
LUCIFERASES; RACEMIC mixtures; MARINE natural products; BIOLUMINESCENCE; HIGH performance liquid chromatography; LUMINESCENCE measurement; CHIRAL centers
- Publication
International Journal of Molecular Sciences, 2024, Vol 25, Issue 5, p2699
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms25052699