10.1021/bi501202u.s001 Bruce R. Branchini Bruce R. Branchini Tara L. Southworth Tara L. Southworth Danielle M. Fontaine Danielle M. Fontaine Audrey L. Davis Audrey L. Davis Curran E. Behney Curran E. Behney Martha H. Murtiashaw Martha H. Murtiashaw A <i>Photinus pyralis</i> and <i>Luciola italica</i> Chimeric Firefly Luciferase Produces Enhanced Bioluminescence American Chemical Society 2014 bioluminescence chimeric enzyme Luciola italica luciferase Luciola italica Chimeric Firefly Luciferase Produces Enhanced BioluminescenceWe report novel PpyLit chimera Photinus pyralis luciferase 2014-10-14 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/A_i_Photinus_pyralis_i_and_i_Luciola_italica_i_Chimeric_Firefly_Luciferase_Produces_Enhanced_Bioluminescence/2245288 We report the enhanced bioluminescence properties of a chimeric enzyme (PpyLit) that contains the N-domain of recombinant <i>Photinus pyralis</i> luciferase joined to the C-domain of recombinant <i>Luciola italica</i> luciferase. Compared to the <i>P. pyralis</i> 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 (<i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub>), 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.