Functional conversion of fatty acyl-CoA synthetase to firefly luciferase by site-directed mutagenesis: A key substitution responsible for luminescence activity |
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Authors: | Yuichi Oba Koichiro Iida |
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Affiliation: | a Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan b Yokohama Research Center, Chisso Co., Kanazawa, Yokohama 236-8605, Japan |
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Abstract: | 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 Drosophilamelanogaster (CG6178) gave luminescence enhancement, suggesting that the serine residue is a key substitution responsible for luminescence activity. |
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Keywords: | CoA, coenzyme A DLSA, 5&rsquo -O-[N-(dehydroluciferyl)-sulfamoyl]adenosine FACS, fatty acyl-CoA synthetase LcLuc, Luciola cruciata luciferase WT, wild-type |
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