Luciferin properties

Properties of luciferin. The crystals are microscopic needles, which melt with decomposition at 205-210°C (Bitler and McElroy, 1957). It is a quite stable luciferin compared with some other luciferins, such as Cypridina luciferin and the luciferins of krill and dinoflagellates. It is not significantly affected by lOmM H2SO4 and lOmM NaOH at room temperature in air. The absorption spectral data of luciferin are shown in Fig. 1.3 (McElroy and Seliger, 1961). The molar absorption coefficient of the 328 nm peak in acidic solutions and that of the 384 nm peak in basic solutions are both 18,200 (Morton et al., 1969). Luciferin is fluorescent, showing an emission maximum at 537 nm in both acidic and basic conditions, although the intensity of the fluorescence is lower in acidic solution than in basic solution (fluorescence quantum yields 0.62 in basic condition, and 0.25 in acidic condition Morton et al., 1969). The chemical synthesis... 

Improved purification method of Cypridina luciferin. The purification of Cypridina luciferin became remarkably simple and easy after some of the properties of this substance were known. The following method was used to obtain the large quantity of luciferin needed for the study of its chemical structure. The method consists of three steps and takes less than eight hours to obtain crystallized luciferin. 

Properties. Cypridina luciferin is soluble in water, methanol and other alcoholic solvents, but not in most aprotic solvents. The ultraviolet absorption spectra of luciferin and oxyluciferin are shown in Fig. 3.1.3. Luciferin in neutral solutions is yellow (lmax 432 nm ... 

Properties of Latia luciferin. Latia luciferin is a highly hydrophobic, fat-soluble compound, and volatile under vacuum. It is a colorless liquid, with an absorption maximum at 207nm (s approx. 13,700 Fig. 6.1.2). The chemical structure of Latia luciferin has been determined to be 1 (C15H24O2), an enol formate of a terpene aldehyde 3 (Fig. 6.1.3 Shimomura and Johnson, 1968b). The enol formate group of Latia luciferin is unstable the luciferin is spontaneously hydrolyzed... 

Properties of luciferin. The luciferin of Odontosyllis is a highly polar substance. It is soluble in water, methanol, and DMF, but practically insoluble in ra-butanol, ethyl acetate and acetonitrile. The luciferin is strongly adsorbed onto DEAE-cellulose, even under acidic conditions, indicating that the molecule possesses a strong acidic functionality. Although the luciferin is unstable in the presence of air, it is quite stable in dilute methanol under argon at — 20°C. 

Structure of luciferin (Ohtsuka et al., 1976). The luciferin of Diplocardia longa is a colorless liquid, and fairly stable at room temperature. It is soluble in polar organic solvents (methanol, ethanol, acetone, and methyl acetate) but insoluble in nonpolar solvents like hexane and carbon tetrachloride. Based on the chemical properties and spectroscopic data, the following chemical structure was assigned to the luciferin. 

The solution of purified dinoflagellate luciferin is yellow, showing absorption maxima at 245 and 390nm in an aqueous solution and at 241 and 388 nm in 40% acetonitrile containing 85 mM NaCl and 3 mM NaHCOs (Fig. 8.4). The compound is strongly fluorescent in blue (excitation maximum at 390 nm, emission maximum at 474 nm Fig. 8.5). The properties of this luciferin are nearly identical with those of the compound F of euphausiid shrimps (Section 3.2). The luciferin is rapidly oxidized in the presence of a trace of oxygen, and also inactivated by a weak acid, even by an acidity of pH 4 or the acidity... 

Properties of luciferin precursors. About one dozen of the luciferin precursors of M. citricolor isolated by HPLC had a strong tendency of isomerization, as mentioned above. Their molecular weights could not be established by mass spectrometry, which is probably due to isomerization, although they appear to be in a range of 300-600. The precursors showed an absorption peak at about 369 nm in methanol and aqueous acetonitrile (Fig. 9.13). According to an NMR study, all precursors probably contain the following common partial structure (personal communication from Dr. H. Nakamura, 1998). 

Properties of the activation product. The two decylamine-activation products (luciferins) showed similar absorption characteristics (A.max 372 nm in water, and 375 nm in ethanol), which clearly differ from the absorption peak of the natural luciferin (320 nm) reported by Kuwabara and Wassink (1966). The fluorescence emission of the activation products varied significantly by solvents, showing a peak at 460 nm in neutral aqueous solution and a broad peak at 485-522 nm in ethanol. They emitted chemiluminescence (A.max 580 nm) in the presence of CTAB, H2O2 and Fe2+ (Fig. 9.13), in resemblance to the (NH4)2S04-activation product of panal (A.max 570 nm). 

Structure determination of luciferin. Once a luciferin is obtained in a sufficient purity, the determination of luciferin structure should be attempted most of the important properties of luciferin are usually already obtained during the course of purification as a necessity. The structural study is considerably more straightforward than the extraction and purification, due to the availability of advanced methods, such as high-resolution mass spectrometry and various NMR techniques. If help or collaboration is needed in structure determination, the attractiveness of a luciferin will make it easy to find a good collaborator. However, the purified luciferin is usually an extremely precious material considering the effort spent in preparing it. To avoid accidental loss of the purified material, the chosen collaborator must have solid knowledge and experience in structure determination a criterion to be considered is that the person has successfully done the structure determination of at least one new natural product. 

Bitler, B., and McElroy, W. D. (1957). The preparation and properties of crystalline firefly luciferin. Arch. Biochem. Biophys. 72 358-368. 

Cormier, M. J., and Charbonneau, H. (1977). Isolation, properties and function of a calcium-triggered luciferin binding protein. In Wasserman, R. H., et al. (eds.), Calcium Binding Proteins and Calcium Function, pp. 481-489. Elsevier North-Holland. 

Henry, J. P., and Michelson, A. M. (1970). Studies in bioluminescence. IV. Properties of luciferin from Pholas dactylus. Biochim. Biophys. Acta 205 451-458. 

Hori, K., and Cormier, M. J. (1966). Studies on the bioluminescence of Renilla reniformis. VI. Some chemical properties and the tentative partial structure of luciferin. Biochim. Biophys. Acta 130 420 125. 

Michelson, A. M. (1978). Purification and properties of Pholas dactylus luciferin and luciferase. Method. Enzymol. 57 385-406. 

Morse, D., Fritz, L., Pappenheimer, A. M., Jr., and Hastings, J. W. (1989). Properties and cellular localization of luciferin binding protein in the bioluminescence reaction of Gonyaulax polyedra. J. Biolumin. Chemi-lumin. 3 79-83. 

Morton, R. A., Hopkins, T. A., and Seliger, H. H. (1969). The spectroscopic properties of firefly luciferin and related compounds. An approach to product emission. Biochemistry 8 1598-1607. 

Rudie, N. G., Ohtsuka, H., and Wampler, J. E. (1976). Purification and properties of luciferin from the bioluminescent earthworm, Diplocardia longa. Photochem. Photobiol. 23 71-73. 

Balanoglossus luciferase, 315 extraction, 315 Balanoglossus luciferin, 316 extraction, 316 Basidiomycetes, 266 Bathophilus, 338 Beneckea, 31, 33, 34, 333 Bentbalbella, 339 Benthosema, 339 2-Benzylimidazopyrazinone, 168 Beroe, 155, 334 Berovin, 155, 346 extraction and purification, 155 spectral properties, 156 BFP, 101... 

The present book describes all the significant studies and findings on the chemistry of the more than 30 different bioluminescent systems presently known, accompanied by over 1000 selected references. It includes descriptions of the purification and properties of bioluminescent compounds, such as luciferins, luciferases and photoproteins, and the mechanisms of luminescence reactions. To make the book more useful than a mere review volume and to save researchers time in looking into original references, I have included a considerable amount of original experimental methods, data and graphs. In addition, I have included some new data and experimental methods unavailable elsewhere. I hope this volume will be useful to researchers and students, and it will be my greatest pleasure if this book contributes... 

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