Random mutagenesis of Luciola mingrelica firefly luciferase. Mutant enzymes with bioluminescence spectra showing low pH sensitivity.
Most firefly luciferases demonstrate a strong pH-dependence of bioluminescence spectra. Gene region encoding first 225 residues of Luciola mingrelica luciferase was subjected to random mutagenesis, and four mutants with altered pH-sensitivity of bioluminescence spectra were isolated. F16L substitution showed distinctly lower pH-dependence of bioluminescence spectra, and Y35N,H and F16L/A40S substitutions resulted in the enzymes with bioluminescence spectra virtually independent from pH in the range of 6.0-7.8. The structural explanation is proposed for the effect of mutations on pH-sensitivity of bioluminescence spectra.
Koksharov
M I
MI
Department of Chemical Enzymology, Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
Ugarova
N N
NN
D000595:Amino Acid Sequence; D000818:Animals; D004795:Enzyme Stability; D048888:Fireflies / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D049409:Luciferases, Firefly / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D049449:Luminescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011487:Protein Conformation; D016415:Sequence Alignment; D013696:Temperature
D049409:Luciferases, Firefly