Random mutagenesis of Luciola mingrelica firefly luciferase. Mutant enzymes with bioluminescence spectra showing low pH sensitivity |
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| Authors: | M. I. Koksharov N. N. Ugarova |
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| Affiliation: | Department of Chemical Enzymology, Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119991, Russia. nugarova@gmail.com. |
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| Abstract: | 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. |
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| Keywords: | firefly luciferase random mutagenesis mutation of amino acid residues bioluminescence spectra pH dependence of bioluminescence spectra |
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