Firefly Luciferin

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Firefly Luciferin Down the Rabbit Hole of Quirality (no Quantum Madness today…) Pathway inside Photinus pyralis Notice that while luciferin is a quiral protein, oxyluciferin is not. How to Recycle oxyluciferin: pathway Luc LRE No one How to Recycle oxyluciferin: enzyme Meet: Luciferine Regenerating Enzyme in Photinus pyralis ergo: • Sustainable reconversion reaction of oxyluciferin into D-luciferin. • Quirality of D-luciferin donated by D-cysteine. • With L-cysteine, oxyluciferin becomes L- luciferine. • However, only D-luciferin has luminescent properties! • Ergo: we need either D-cysteine or D-luciferin Enter Magic Enzyme • Magic Enzyme will stereoisomericaly bio- inverse L-luciferin into D-luciferin • Magic Enzyme will be simple and cheap • Magic Enzyme will be coded by a few genes • Magic Enzyme will be expressable in E. coli • Magic Enzyme won’t have complexities… • Does Magic Enzyme exist? A Voice from Beyond • “We previously reported that in the presence of ATP, Mg2+ and CoA, firefly luciferase exhibits coenzyme A ligase (CoA-ligase) activity on L-luciferin in vitro, but not on D- luciferin, to give luciferyl-CoA” Magic Quirality Conversion The Prestige • Magic enzyme is Luciferase! • “Lembert first reported the light production from L-luciferin and proposed that L-luciferin was racemized to give D-luciferin and it was effectively stimulated by the addition of pyrophosphate. This racemization is an in vitro reaction of luciferase in the absence of CoA. In the presence of CoA, L-luciferin is readily converted into luciferyl-CoA.” Huh?! • Initial medium inoculation with D-luciferin • Luciferase takes D-luciferin, produces oxyluciferin • Oxyluciferin is tranformed into L-luciferin via action of LRE with L-cysteine • L-luciferin is transformed into D-luciferin through Luciferase action So: • We CAN use Photinus pyralis luciferase • Suggested red-shifted mutant: Ser284Thr shines at 615nm Sources.

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William D. Mcelroy Papers
http://oac.cdlib.org/findaid/ark:/13030/kt6489r0v5 No online items William D. McElroy Papers Special Collections & Archives, UC San Diego Special Collections & Archives, UC San Diego Copyright 2005 9500 Gilman Drive La Jolla 92093-0175 [email protected] URL: http://libraries.ucsd.edu/collections/sca/index.html William D. McElroy Papers MSS 0483 1 Descriptive Summary Languages: English Contributing Institution: Special Collections & Archives, UC San Diego 9500 Gilman Drive La Jolla 92093-0175 Title: William D. McElroy Papers Identifier/Call Number: MSS 0483 Physical Description: 10.4 Linear feet(19 archives boxes, 11 oversize folders, 6 art bin items) Date (inclusive): 1944-1999 Abstract: Papers of William David McElroy (1917-1999), professor of biochemistry, the fourth chancellor (1972-1980) of the University of California, San Diego; and former director (1969-1971) of the National Science Foundation. Scope and Content of Collection Papers of William David McElroy (1917-1999), professor of biochemistry, the fourth chancellor (1972-1980) of the University of California, San Diego; and former director (1969-1971) of the National Science Foundation. McElroy's significant contributions to biology include isolating and crystallizing the compounds that enable firefly luminescence and for his subsequent research into bacterial bioluminescence. He also wrote, spoke, and worked on problems in areas of environment, pollution, food production, science education, and international science. The papers largely document McElroy's scientific research and include correspondence with the scientific community, various biographical materials including awards and photographs, his trip to China in 1979, writings and reprints related to biochemical and scientific investigation, research materials on bioluminescence, teaching materials, and speeches given both as chancellor and as director of the National Science Foundation.
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Safety Data Sheet
G-Biosciences, St Louis, MO, USA | 1-800-628-7730 | 1-314-991-6034 | [email protected] A Geno Technology, Inc. (USA) brand name Safety Data Sheet Cat. # RC-227 D-Luciferin Firefly, FREE ACID Size: 0.25g think proteins! think G-Biosciences! www.GBiosciences.com D-Luciferin Firefly, potassium salt Safety Data Sheet according to Federal Register / Vol. 77, No. 58 / Monday, March 26, 2012 / Rules and Regulations Date of issue: 05/06/2013 Revision date: 05/11/2017 Version: 7.1 SECTION 1: Identification 1.1. Identification Product form : Substance Trade name : D-Luciferin Firefly, potassium salt CAS-No. : 2591-17-5 Product code : 006A Formula : C11H8N2O3S2 Synonyms : (4S)-4,5-dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid / (S)-2-(6-hydroxy-2- benzothiazolyl)-2-thiazoline-4-carboxylic acid / (S)-4,5-dihydro-2-(6-hydroxybenzothiazol-2- yl)thiazole-4-carboxylic acid / 2-(6-hydroxy-2-benzothiazolyl)-2-thiazoline-4-carcoxylic acid, (S)- / 2-(6-hydroxybenzothiazol-2-yl)-2-thiazoline-4-carboxylic acid / 4,5-dihydro-2-(6-hydroxy-2- benzorhiazolyl)-4-thiazolecarboxylic acid, (4S)- / 4,5-dihydro-2-(6-hydroxy-2-benzothiazolyl)-4- thiazolecarboxylic acid / 4-Thiazolecarboxylic acid, 4,5-dihydro-2-(6-hydroxy-2-benzothiazolyl)-, (S)- / D-(-)-luciferin / D-luciferin / firefly luciferin / liciferin, D-(-)- / luciferin / luciferin, D- Other means of identification : D-Luciferin Firefly, free acid 4,5-Dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid, ST50405784, Luciferin, CHEBI:17165 BIG No : 48631 1.2. Recommended use and restrictions on use Use of the substance/mixture : Luciferin is a common bioluminescent reporter used for in-vivo imaging of the expression of the luc marker gene .
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Bioluminescence Is Produced by a Firefly-Like Luciferase but an Entirely
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(12) United States Patent (10) Patent No.: US 9,574.223 B2 Cali Et Al
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Presentation1
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Oxidation of Firefly Luciferin (Luminescence/Luciferase/Oxygen Exchange/Reaction Mechanism) OSAMU SHIMOMURA*, TOSHIO Gotot, and FRANK H
Proc. Nati. Acad. Scf. USA Vol. 74, No. 7, pp. 2799-2802, July 1977 Biochemistry Source of oxygen in the CO2 produced in the bioluminescent oxidation of firefly luciferin (luminescence/luciferase/oxygen exchange/reaction mechanism) OSAMU SHIMOMURA*, TOSHIO GOTOt, AND FRANK H. JOHNSON* * Department of Biology, Princeton University, Princeton, New Jersey 08540; and t Department of Agricultural Chemistry, Nagoya University, Chikusa, Nagoya 464, Japan Communicated by Henry Eyring, May 2, 1977 ABSTRACT Incorporation of 180 into the CO2 produced in luciferin by labeling product CO2 with 180 fully supported the bioluminescent oxidation of firefly luciferin was studied. scheme 1 (13-15). In regard to Renilla luciferin, however, one In H2160 medium with 1802 gas, the product CO2 contained up 0 of the was to arise from solvent to 75% C'60180, showing that one 0 of the product CO2 arose product CO2 reported H120 from the 02 that oxidized luciferin. This result is consistent with in both bioluminescence (3) and chemiluminescence (12), a dioxetane mechanism. Analysis of the mass spectral data of which would seem odd in view of the structural similarity be- the CO2 obtained in high-enrichment H2180 medium with 1602 tween Renilla luciferin and Cypridina luciferin. gas indicated the presence of about 20% contaminating CO2, The present study unambiguously supports scheme 1, but not which contributes approximately 70% of the total incorporated scheme 2, for the bioluminescence of firefly luciferin. More- 180. Thus the values of incorporated 180 in 112180 medium with over, present data cast a serious doubt on the validity of pre- 1802 gas have no significance in the present context.
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Illuminating Insights Into Firefly Luciferase and Other
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