Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase
Cypridina luciferin (CypL) is a marine natural product that functions as the luminous substrate for the enzyme <i>Cypridina</i> luciferase (CypLase). CypL has two enantiomers, (<i>R</i>)- and (<i>S</i>)-CypL, due to its one chiral center at the <i>sec</i&...
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/5/2699 |
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| author | Shusei Kanie Chun Wu Kiyohito Kihira Rie Yasuno Yasuo Mitani Yoshihiro Ohmiya |
| author_facet | Shusei Kanie Chun Wu Kiyohito Kihira Rie Yasuno Yasuo Mitani Yoshihiro Ohmiya |
| author_sort | Shusei Kanie |
| collection | DOAJ |
| description | Cypridina luciferin (CypL) is a marine natural product that functions as the luminous substrate for the enzyme <i>Cypridina</i> luciferase (CypLase). CypL has two enantiomers, (<i>R</i>)- and (<i>S</i>)-CypL, due to its one chiral center at the <i>sec</i>-butyl moiety. Previous studies reported that (<i>S</i>)-CypL or racemic CypL with CypLase produced light, but the luminescence of (<i>R</i>)-CypL with CypLase has not been investigated. Here, we examined the luminescence of (<i>R</i>)-CypL, which had undergone chiral separation from the enantiomeric mixture, with a recombinant CypLase. Our luminescence measurements demonstrated that (<i>R</i>)-CypL with CypLase produced light, indicating that (<i>R</i>)-CypL must be considered as the luminous substrate for CypLase, as in the case of (<i>S</i>)-CypL, rather than a competitive inhibitor for CypLase. Additionally, we found that the maximum luminescence intensity from the reaction of (<i>R</i>)-CypL with CypLase was approximately 10 fold lower than that of (<i>S</i>)-CypL with CypLase, but our kinetic analysis of CypLase showed that the K<sub>m</sub> value of CypLase for (<i>R</i>)-CypL was approximately 3 fold lower than that for (<i>S</i>)-CypL. Furthermore, the chiral high-performance liquid chromatography (HPLC) analysis of the reaction mixture of racemic CypL with CypLase showed that (<i>R</i>)-CypL was consumed more slowly than (<i>S</i>)-CypL. These results indicate that the turnover rate of CypLase for (<i>R</i>)-CypL was lower than that for (<i>S</i>)-CypL, which caused the less efficient luminescence of (<i>R</i>)-CypL with CypLase. |
| first_indexed | 2024-04-25T00:29:08Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-04-25T00:29:08Z |
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| spelling | doaj.art-8a23563c33a94254a51313ac68aff30d2024-03-12T16:46:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01255269910.3390/ijms25052699Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> LuciferaseShusei Kanie0Chun Wu1Kiyohito Kihira2Rie Yasuno3Yasuo Mitani4Yoshihiro Ohmiya5Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hokkaido Center, 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, JapanBiomedical Research Institute, AIST, Kansai Center, 1-8-31 Midorigaoka, Ikeda 563-8577, JapanJapan Aerospace Exploration Agency (JAXA), Tsukuba Space Center, 2-1-1 Sengen, Tsukuba 305-8505, JapanCellular and Molecular Biotechnology Research Institute, AIST, Tsukuba Center, 1-1-1 Higashi, Tsukuba 305-8566, JapanBioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hokkaido Center, 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, JapanBiomedical Research Institute, AIST, Kansai Center, 1-8-31 Midorigaoka, Ikeda 563-8577, JapanCypridina luciferin (CypL) is a marine natural product that functions as the luminous substrate for the enzyme <i>Cypridina</i> luciferase (CypLase). CypL has two enantiomers, (<i>R</i>)- and (<i>S</i>)-CypL, due to its one chiral center at the <i>sec</i>-butyl moiety. Previous studies reported that (<i>S</i>)-CypL or racemic CypL with CypLase produced light, but the luminescence of (<i>R</i>)-CypL with CypLase has not been investigated. Here, we examined the luminescence of (<i>R</i>)-CypL, which had undergone chiral separation from the enantiomeric mixture, with a recombinant CypLase. Our luminescence measurements demonstrated that (<i>R</i>)-CypL with CypLase produced light, indicating that (<i>R</i>)-CypL must be considered as the luminous substrate for CypLase, as in the case of (<i>S</i>)-CypL, rather than a competitive inhibitor for CypLase. Additionally, we found that the maximum luminescence intensity from the reaction of (<i>R</i>)-CypL with CypLase was approximately 10 fold lower than that of (<i>S</i>)-CypL with CypLase, but our kinetic analysis of CypLase showed that the K<sub>m</sub> value of CypLase for (<i>R</i>)-CypL was approximately 3 fold lower than that for (<i>S</i>)-CypL. Furthermore, the chiral high-performance liquid chromatography (HPLC) analysis of the reaction mixture of racemic CypL with CypLase showed that (<i>R</i>)-CypL was consumed more slowly than (<i>S</i>)-CypL. These results indicate that the turnover rate of CypLase for (<i>R</i>)-CypL was lower than that for (<i>S</i>)-CypL, which caused the less efficient luminescence of (<i>R</i>)-CypL with CypLase.https://www.mdpi.com/1422-0067/25/5/2699Cypridina luciferinbioluminescencemarine natural product<i>Cypridina</i> luciferasechiral separation |
| spellingShingle | Shusei Kanie Chun Wu Kiyohito Kihira Rie Yasuno Yasuo Mitani Yoshihiro Ohmiya Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase International Journal of Molecular Sciences Cypridina luciferin bioluminescence marine natural product <i>Cypridina</i> luciferase chiral separation |
| title | Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase |
| title_full | Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase |
| title_fullStr | Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase |
| title_full_unstemmed | Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase |
| title_short | Bioluminescence of (<i>R</i>)-Cypridina Luciferin with <i>Cypridina</i> Luciferase |
| title_sort | bioluminescence of i r i cypridina luciferin with i cypridina i luciferase |
| topic | Cypridina luciferin bioluminescence marine natural product <i>Cypridina</i> luciferase chiral separation |
| url | https://www.mdpi.com/1422-0067/25/5/2699 |
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