SUPPORTING INFORMATION Structure and Spectroscopy of Oxyluciferin, the Light Emitter of the Firefly Bioluminescence Panče Naumov,* Yutaka Ozawa, Kei Ohkubo, and Shunichi Fukuzumi Department of Material and Life Science, and Frontier Research Center for Global Young Researchers, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka, Japan
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Figure S1. Solid-state absorption (top) and emission (bottom, ex = 400 nm) of oxyluciferin and 5-methyloxyluciferin in the solid state.
Figure S2. 1H NMR spectrum of oxyluciferin in acetonitrile-d3.
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Figure S3. 1H NMR spectrum of oxyluciferin in methanol-d4.
Figure S4. Temporal profile (A) and effect of the excess base on the absorption spectra of 5methyloxyluciferin and its deprotonated forms in DMSO under anaerobic conditions. The numbers in the legend of panel A are the respective times (in minutes) from the mixing of the reactants (“1” stands for solution before the addition of base). The numbers in the legend of panel B are the approximate relative ratios of the base. The numbers represent the band maximum wavelengths in nanometers. S3
Figure S5. 1H NMR spectrum of oxyluciferin in DMSO-d6
Figure S6. 1H NMR spectrum of 5-methyloxyluciferin in DMSO-d6 S4
Table S1. Crystallographic data for oxyluciferin and 5-methyloxyluciferin oxyluciferin C10H6N2O2S2 250.29 293 0.71073 Monoclinic, P21/n a = 4.8806(4) Å b = 18.8242(14) Å c = 11.1298(8) Å = 90o = 99.260(5)o = 90 deg. 1009.21(13) 4, 1.647 0.510 512 0.31 × 0.10 × 0.07 2.15 to 26.39
5-methyloxyluciferin C11H8N2O2S2 264.31 293 0.71073 Monoclinic, P21/n a = 6.3027(4) Å b = 14.8046(10) Å c = 11.6515(7) Å = 90o = 91.419(4)o = 90o 1086.86(12) 4, 1.615 0.478 544 0.22 × 0.03 × 0.03 2.22 to 27.48
Limiting indices
5 ← h ←6, 20 ← k ←23 13 ← l ←13
8 ← h ←8 19 ← k ←14 15 ← l ←14
Reflections collected / unique Completeness / % Max. and min. transmission Refinement method Data / restraints / parameters Goodness-of-fit on F2 Final R indices [I > 2(I)] R indices (all data) Largest diff. peak and hole
14599 / 2056 (Rint = 0.0554) 99.6 0.9676 and 0.8579 FMLS 2056 / 0 / 157 1.037 R1 = 0.0524, wR2 =0.1239 R1 = 0.0913, wR2 =0.1412 0.327 and 0.252 e Å3
9595 / 2476 (Rint = 0.0261) 99.5 0.9858 and 0.9021 FMLS 2476 / 0 / 163 1.069 R1 = 0.0337, wR2 = 0,0875 R1 = 0.0546, wR2 = 0.0984 0.350 and 0.200 e Å3
Empirical formula Formula weight Temperature / K Wavelength / Å Crystal system, space group Unit cell dimensions
Volume / Å3 Z, Calc. density / Mg m3 Absorption coeff. / mm1 F(000) Crystal size / mm Theta range / o
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Complete reference 66: GAUSSIAN03, Revision B.05, M. J. Frisch, G. W. Trucks, H. B. Schlegel, G. E. Scuseria, M. A. Robb, J. R. Cheeseman, J. A. Montgomery, Jr., T. Vreven, K. N. Kudin, J. C. Burant, J. M. Millam, S. S. Iyengar, J. Tomasi, V. Barone, B. Mennucci, M. Cossi, G. Scalmani, N. Rega, G. A. Petersson, H. Nakatsuji, M. Hada, M. Ehara, K. Toyota, R. Fukuda, J. Hasegawa, M. Ishida, T. Nakajima, Y. Honda, O. Kitao, H. Nakai, M. Klene, X. Li, J. E. Knox, H. P. Hratchian, J. B. Cross, V. Bakken, C. Adamo, J. Jaramillo, R. Gomperts, R. E. Stratmann, O. Yazyev, A. J. Austin, R. Cammi, C. Pomelli, J. W. Ochterski, P. Y. Ayala, K. Morokuma, G. A. Voth, P. Salvador, J. J. Dannenberg, V. G. Zakrzewski, S. Dapprich, A. D. Daniels, M. C. Strain, O. Farkas, D. K. Malick, A. D. Rabuck, K. Raghavachari, J. B. Foresman, J. V. Ortiz, Q. Cui, A. G. Baboul, S. Clifford, J. Cioslowski, B. B. Stefanov, G. Liu, A. Liashenko, P. Piskorz, I. Komaromi, R. L. Martin, D. J. Fox, T. Keith, M. A. Al-Laham, C. Y. Peng, A. Nanayakkara, M. Challacombe, P. M. W. Gill, B. Johnson, W. Chen, M. W. Wong, C. Gonzalez, and J. A. Pople, Gaussian, Inc., Wallingford CT, 2004.
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