Biosynthesis-inspired deracemizative production of d-luciferin by combining luciferase and thioesterase
| Autor: | Kazunari Arima, Kazuki Niwa, Masatoshi Okuda, Masahiro Takeo, Juri Maeda, Yuji Ito, Dai-ichiro Kato, Seiji Negoro |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Luminescence Biophysics Firefly Luciferin Biology Biochemistry 03 medical and health sciences chemistry.chemical_compound Biosynthesis Thioesterase Animals Bioluminescence Luciferase Luciferases Molecular Biology chemistry.chemical_classification 030102 biochemistry & molecular biology Fireflies Substrate (chemistry) Stereoisomerism Luciferin 030104 developmental biology Enzyme Palmitoyl-CoA Hydrolase chemistry Light emission |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - General Subjects. 1861:2112-2118 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/j.bbagen.2017.04.010 |
| Popis: | Due to the strict enantioselectivity of firefly luciferase, only d-luciferin can be used as a substrate for bioluminescence reactions. Unfortunately, luciferin racemizes easily and accumulation of nonluminous l-luciferin has negative influences on the light emitting reaction. Thus, maintaining the enantiopurity of luciferin in the reaction mixture is one of the most important demands in bioluminescence applications using firefly luciferase. In fireflies, however, l-luciferin is the biosynthetic precursor of d-luciferin, which is produced from the L-form undergoing deracemization. This deracemization consists of three successive reactions: l-enantioselective thioesterification by luciferase, in situ epimerization, and hydrolysis by thioesterase. In this work, we introduce a deracemizative luminescence system inspired by the biosynthetic pathway of d-luciferin using a combination of firefly luciferase from Luciola cruciata (LUC-G) and fatty acyl-CoA thioesterase II from Escherichia coli (TESB). The enzymatic reaction property analysis indicated the importance of the concentration balance between LUC-G and TESB for efficient d-luciferin production and light emission. Using this deracemizative luminescence system, a highly sensitive quantitative analysis method for l-cysteine was constructed. This LUC-G-TESB combination system can improve bioanalysis applications using the firefly bioluminescence reaction by efficient deracemization of D-luciferin. |
| Databáze: | OpenAIRE |
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