Luciola mingrelica

Devine, J. H., et al. (1993). Luciferase from the east European firefly Luciola mingrelica cloning and nucleotide sequence of the cDNA overexpression in Escherichia coli and purification of enzyme. Biochim. Bio-phys. Acta 1173 121-132. 

Although numerous luminous organisms are known, only a few of them has been studied and really exploited. Analytical applications of bioluminescence concern mainly the detection of ATP with the firefly luciferase and of NADH with some marine bacteria systems. Luciferase from the North American firefly, i.e., Photinus pyralis, has been extensively studied10-12 and afterwards, attention has been paid to the luciferase from Luciola mingrelica, i.e., the North Caucasus firefly13 15. 

Ugarova N.N., Luciferase of Luciola mingrelica fireflies. Kinetics and regulation mechanism, J. Biolumin. Chemilumin. 1989 4 406-418. 

Beside that of Photinus pyralis, other luciferases have been described in the literature such as those from Luciola cruciata, Luciola lateralis [122], and Luciola mingrelica. Among them luciferase from the latter one has been employed for analytical purposes [123],... 

In this age of powerful computers, it is no longer even necessary to find analytical solution to differential equations. There are many software packages available that cany out numerical integration of differential equations followed by non-linear regression to fit the model and assess its quality by comparing with experimental data. In this study we have used a numerical integration approach to compare kinetic properties of Photinus pyralis and Luciola mingrelica firefly luciferases. 

Photinus pyralis firefly luciferase (PP) was obtained from Sigma, Luciola mingrelica firefly luciferase (LM) was isolated and purified according to. Time-course of bioluminescent reaction rate (v) was monitored as the intensity of light (I) in time according to equation 1 using a luminometer model 1251 (LKB Sweden). 

Dementieva El, Kutizova GD, Ugarova NN. Biochemical properties and stability of homogeneous luciferase of fireflies Luciola mingrelica. Vestn Mosk U Khim 1989 30 601-6. 

Leontieva O, Vlasova T, Ugarova N. Interaction of firefly lucifaase Luciola mingrelica with dimethyloxyluciferin. In Stanley P, Kricka L. eds. Bioluminescence Chemiluminescence. Progress and Current Applications. Singapore World Scientific. 2002 41-4. 

Luminometer 3550i and Filtravette (New Horizons Diagnostics Corp., USA) were used for bioluminescent measurement. ATP-reagent (based on soluble Luciola mingrelica firefly luciferase) was developed in our laboratory. Dimethyl sulfoxide (DMSO) was from Reakhim (Russia). Nutrition broth (NB) from ICN, Tryptic Soy Broth (TSB) from Difco and Thioglycolate broth (TB) from Merck were used. Ultrapure deionized water was obtained on Milli-Q (Millipore). Broth culture of Escherichia coli LE392 was used in model experiments. 

ENHANCEMENT OF THERMOSTABILITY OF LUCIOLA MINGRELICA FIREFLY LUCIFERASE BY MUTAGENESIS OF NON-CONSERVATIVE RESIDUES CYS62 AND CYS146... 

Table 1. Kinetic rate constants of thermal inactivation for the WT firefly luciferase Luciola mingrelica and its mutant forms at 37°C at the enzyme concentration 106 - 108mol/L. Conditions 0.05 mol/L Tris-acetate buffer,...

Fig. 1. Time-courses of enzyme inactivation at 37°C of WT Luciola mingrelica luciferase and its mutant forms with point mutations Cys62Ser and Cysl46Ser in the absence (filled triangles) and presence (open circles) of 12 mmol/L dithiothreitol. The enzyme concentration was 10 7 mol/L. For other reaction conditions, please...

Dementieva E, Kutuzova G, Zheleznova E, Lundovskikh 1, Ugarova N. Physicochemical properties of recombinant Luciola mingrelica luciferase and its mutant forms. Biochemistry (Moscow) 1996 61 115-9. 

Brovko L, Ugarova N, Kinetics and mechanism of inactivation and reactivation of immobilized luciferase from fireflies Luciola mingrelica and the role of sulfhydryl in these processes. Biokhimiia (in Russian) 1980 45 794-801. 

Lundovskikh I, Dementieva E, Ugarova N, Kinetics and mechanism of thermo inactivation of firefly luciferase Luciola mingrelica. Moscow Univ Chem Bull 2000 41 362-6. 

The majority of firefly luciferases demonstrate a pH-dependence of bioluminescence spectra. Analyzing bioluminescence spectra over a large pH interval, the relative contribution of different (LO ) forms to the total bioluminescence spectra of native and some mutant luciferases was characterized quantitatively.2 9 For example, the point mutation H433Y in Luciola mingrelica firefly luciferase resulted in 40 nm... 

Koksharov M, Ugarova N. Random mutagenesis of Luciola mingrelica firefly luciferase. Mutant enzymes whose bioluminescence spectra show low pH-sensitivity. Biochemistry (Moscow) 2008 73 in press. 

M Tricine-NaOH, pH 8.4. ATP was determined by bioluminescence using the immobilized luciferase from Luciola mingrelica (8). The yield ATP amounted to about 2% of the initial ADP. 

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