title: A Photinus pyralis and Luciola italica chimeric firefly luciferase produces enhanced bioluminescence.
abstract: We report the enhanced bioluminescence properties of a chimeric enzyme (PpyLit) that contains the N-domain of recombinant Photinus pyralis luciferase joined to the C-domain of recombinant Luciola italica luciferase. Compared to the P. pyralis enzyme, the novel PpyLit chimera exhibited 1.8-fold enhanced flash-height specific activity, 2.0-fold enhanced integration-based specific activity, 2.9-fold enhanced catalytic efficiency (kcat/Km), and a 1.4-fold greater bioluminescence quantum yield. The results of this study provide an underlying basis of this unusual example of a chimeric enzyme with enhanced catalytic properties that are not simply the sum of the contributions of the two luciferases.
authors.affiliation: Department of Chemistry, Connecticut College , New London, Connecticut 06320, United States.
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Branchini
authors.forename: Tara L
authors.initials: TL
authors.lastname: Southworth
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Fontaine
authors.forename: Audrey L
authors.initials: AL
authors.lastname: Davis
authors.forename: Curran E
authors.initials: CE
authors.lastname: Behney
authors.forename: Martha H
authors.initials: MH
authors.lastname: Murtiashaw
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D057075:Enzyme Assays; D048888:Fireflies / Q000201:enzymology*; D049454:Firefly Luciferin / Q000737:chemistry; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008163:Luminescent Measurements; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly