The enzyme, which is found in fireflies (Lampyridae), is responsible for their biolouminescence. The reaction begins with the formation of an acid anhydride between the carboxylic group of D-firefly luciferin and AMP, with the release of diphosphate. An oxygenation follows, with release of the AMP group and formation of a very short-lived peroxide that cyclizes into a dioxetanone structure, which collapses, releasing a CO2 molecule. The spontaneous breakdown of the dioxetanone (rather than the hydrolysis of the adenylate) releases the energy (about 50 kcal/mole) that is necessary to generate the excited state of oxyluciferin. The excited luciferin then emits a photon, returning to its ground state. The enzyme has a secondary acyl-CoA ligase activity when acting on L-firefly luciferin (see EC 6.2.1.52).
The enzyme, which is found in fireflies (Lampyridae), is responsible for their biolouminescence. The reaction begins with the formation of an acid anhydride between the carboxylic group of D-firefly luciferin and AMP, with the release of diphosphate. An oxygenation follows, with release of the AMP group and formation of a very short-lived peroxide that cyclizes into a dioxetanone structure, which collapses, releasing a CO2 molecule. The spontaneous breakdown of the dioxetanone (rather than the hydrolysis of the adenylate) releases the energy (about 50 kcal/mole) that is necessary to generate the excited state of oxyluciferin. The excited luciferin then emits a photon, returning to its ground state. The enzyme has a secondary acyl-CoA ligase activity when acting on L-firefly luciferin (see EC 6.2.1.52).
construction of a chimeric enzyme luc2 that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase, the recombinant chimeric enzyme shows 2fold enhanced activity and 1.4fold greater bioluminescence quantum yield compared to the wild-type Photinus pyralis luciferase. Further engineering to enhance thermal and pH stability produces a different luciferase called PLG2, that shows 4.4fold enhanced activity and 2.2fold greater bioluminescence quantum yield compared to the wild-type. Five amino acid changes based on Luciola italica are the main determinants of the improved bioluminescence properties
construction of a chimeric enzyme luc2 that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase, the recombinant chimeric enzyme shows 2fold enhanced activity and 1.4fold greater bioluminescence quantum yield compared to the wild-type Photinus pyralis luciferase. Further engineering to enhance thermal and pH stability produces a different luciferase called PLG2, that shows 4.4fold enhanced activity and 2.2fold greater bioluminescence quantum yield compared to the wild-type. Five amino acid changes based on Luciola italica are the main determinants of the improved bioluminescence properties
construction of a chimeric enzyme PpyLit that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase, the recombinant chimeric enzyme PpyLit shows 1.8fold enhanced flash-height specific activity, 2.0fold enhanced integration-based specific activity, 2.9fold enhanced catalytic efficiency (kcat/Km), and a 1.4fold greater bioluminescence quantum yield compared to the wild-type Photinus pyralis luciferase. A chimeric enzyme with enhanced catalytic properties that are not simply the sum of the contributions of the two luciferases
construction of a chimeric enzyme PpyLit that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase, the recombinant chimeric enzyme PpyLit shows 1.8fold enhanced flash-height specific activity, 2.0fold enhanced integration-based specific activity, 2.9fold enhanced catalytic efficiency (kcat/Km), and a 1.4fold greater bioluminescence quantum yield compared to the wild-type Photinus pyralis luciferase. A chimeric enzyme with enhanced catalytic properties that are not simply the sum of the contributions of the two luciferases
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant expression of the chimeric mutant luc2 that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase in HEK-293 cells and in Escherichia coli strain BL21(DE3)pLysS