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Functional conversion of fatty acyl‐CoA synthetase to firefly luciferase by site‐directed mutagenesis: A key substitution responsible for luminescence activity
Author(s) -
Oba Yuichi,
Iida Koichiro,
Inouye Satoshi
Publication year - 2009
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2009.05.018
Subject(s) - luciferase , bioluminescence , luciferases , serine , biochemistry , luciferin , serine hydroxymethyltransferase , mutagenesis , chemistry , drosophila melanogaster , biology , enzyme , mutant , gene , transfection
We demonstrated that firefly luciferase has a catalytic function of fatty acyl‐CoA synthesis [Oba, Y., Ojika, M. and Inouye, S. (2003) Firefly luciferase is a bifunctional enzyme: ATP‐dependent monooxygenase and a long chain fatty acyl‐CoA synthetase. FEBS Lett. 540, 251–254] and proposed that the evolutionary origin of beetle luciferase is a fatty acyl‐CoA synthetase (FACS) in insect. In this study, we performed the functional conversion of FACS to luciferase by replacing a single amino acid to serine. This serine residue is conserved in luciferases and possibly interacts with luciferin. The mutants of FACSs in non‐luminous click beetle Agrypnus binodulus (AbLL) and Drosophila melanogaster (CG6178) gave luminescence enhancement, suggesting that the serine residue is a key substitution responsible for luminescence activity.