CATALYTIC PROPERTIES AND BIOLUMINESCENCE SPECTRA OF RECOMBINANT FIREFLY LUCIFERASE LUCIOLA MINGRELICA WITH POINT MUTATIONS OUT OF THE ENZYME ACTIVE SITE

摘要

The His433Tyr mutation in Luciola cruciata and Hotaria parvula firefly luciferases resulted in bioluminescence color changes from 570 to 610 nm. It means that the highly conservative His433 residue located out of the luciferase active site plays a very important role in enzyme function. The goal of this work was to construct mutants His433Asn and His433Ser for Luciola mingrelica firefly luciferase, which has high homology with luciferases indicated above. We studied catalytic properties of the enzyme mutant forms and their bioluminescence spectra. Analysis of the data obtained permitted us to elucidate the mechanism of the influence of the His433 residue on the luciferase active site.