Luciola

Later, fireflv oxyluciferin was successfully synthesi2ed (403. 408) and has been isolated and identified in firefly lanterns (luciola cruaciata) after the lanterns were treated with pyridine and acetic anhydride to prevent decomposition (409). In 1972, Suzuki and Goto firmly established that oxyluciferin is involved in the bioluminescence of firefly lanterns and in the chemiluminescence of firefly luciferin (403. 410).. A. mechanism involving a four-membered ring cyclic peroxide has been proposed for the reaction (406. 411). However, it was not confirmed by 0 -labelinE experiments (412). 

Fig. 1.1 The firefly Luciola cruciata (male) drawn by Sakyo Kanda (1874-1939), a pioneer of the study of bioluminesccnce in Japan, showing his extraordinary artistic talent (reproduced from Kanda, 1935).

Lampyridae Lampyris, Photinus, Photuris, Luciola, Aspisoma, etc. 

Dement eva, E. I., et al. (1986). The pH-dependence of the bioluminescence spectra and kinetic constant for luciferase of the firefly Luciola mingre-lica. Biokhimiya 51 130-139. 

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. 

Kishi, Y., et al. (1968). Luciferin and luciopterin isolated from the Japanese firefly, Luciola cruciata. Tetrahedron Lett., pp. 2847-2850. 

Ohmiya, Y., Ohba, N., Toh, H., and Tsuji, F. I. (1994). Comparative aspects of the bioluminescence reactions of Hotaria parvula, Luciola cruciata, and Pbotinus pyralis. In Campbell, A. K., etal. (eds.), Biolumin. Chemi-lumin. Proc. Int. Symp., 8th 1994, pp. 572-575. Wiley, Chichester, UK. 

Tatsumi, H., Masuda, T., Kajiyama, N., and Nakano, E. (1989). Luciferase cDNA from Japanese firefly, Luciola cruciata cloning, structure and expression in Escherichia coli. J. Biolumin. Chemilumin. 3 75-78. 

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],... 

COMPARISON OF KINETIC PROPERTIES OF FIREFLY LUCIFERASE FROM PHOTINUS PYRALIS AND LUCIOLA MINGREUCA... 

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. 

Luciola cruciata specimens were collected from 62 sites in Japan covering almost all the insect s distribution areas. Luciola lateralis from 46 sites in Japan and two sites in Korea. Luciola owadai fi om Kume-jima Island, the Okinawa Islands, Japan. 

Figure 1. Distribution pattern of CO II haplotype-groups in Luciola cruciata.

Ohba N. Synchronous flashing in the Japanese firefly, Luciola cruciata (Coleoptera Lampyridae). Sci Rept Yokosuka City Mus 1984 32 23-32. 

Ohba N, Tsumuraya T, Honda K, et al. Ecological study of the firefly, Luciola lateralis, of the Kushiro Shitsugen and Akkeshi, Hokkaido. Sci Rept Yokosuka City Mus 1993 41 15-26. 

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. 

The problem of a poor detection limit was caused by high background ATP and by the low sensitivity of the luciferin-luciferase (L-L) reagent. We have already developed an ATP elimination system using two ATP degrading enzymes (adenosine phosphate deaminase and apyrase) and a surfactant tolerant luciferase that was a mutated Luciola lateralis firefly luciferase. We optimized this elimination system, and investigated its suitability as a detection system. 

PPDK from Microbispora rosea subsp. Aerata (EC2.7.9.1) and thermostable Luciola cruciata firefly luciferase (EC 1.13.12.7) were obtained from Kikkoman Co. (Chiba, Japan). Pyrophosphate, Lysozyme and Proteinase K were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). dNTP Mixture, Taq DNA Polymerase and lOx PCR buffer were acquired from Takara Shuzo Co., Ltd (Osaka, Japan). Primers were synthesized by Takara Shuzo Co., Ltd (Osaka, Japan). Luciferin was obtained from Sigma Chemical Co (St. Louis, MO). Perfect match (PCR Enhancer) was manufactured by Toyobo. Other reagents were of analytical grade. 

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