発光生物学研究室

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Luminous Organisms Lab

Eco-Evo Bioluminescence

We are studying on BIOLUMINESCENCE. From DNA barcoding, molecular phylogeny, biological function, to biosynthesis of luciferins, molecular identification of luciferases, evolution of bioluminescence.

LLL (Living Lights List)

We are listing bioluminescent organisms in Japan and also in the world. Please see the following LLL site.

別サイトにリンクしますhttps://www3.chubu.ac.jp/faculty/oba_yuichi/Living%20Light%20List/

Staff

Yuichi OBA Dr., Associate Professor, PI

Members

Masashi Naito Mr. (DC2)

Noriomi Fujimori Mr. (DC2)

Jose Paitio Mr. (DC2)

Ikuhiko Kin (DC1)

Manabu Bessho (Doctral Researcher)

Hidehiko Hirata (Researcher)

Kaede Yamanaka (BC4)

Yutaro Kuwahara (BC4)

Hidenobu Koike (BC4)

Yusuke Mizutani (BC4)

Tomohiro Mohri (BC4)

Ryohei Inagaki (BC4)

Yuya Kato (BC4)

Hiroki Ajioka (BC4)

Hiroki Sato (BC4)

Shunpei Ukita (BC4)

Our Researches

ALL about BIOLUMINESCENCE!

Luminescent animals occur in 11 phyla and their luminescence systems have evolved independently. So then what is the origin of “Luciferase” enzyme? Where “Luciferin” substrate comes from? The evolutionary process for their curious chemistries remains largely unknown (1, 2). I am focusing on the biochemistry of luminescent animals and its evolutionary aspects using biochemistry, molecular biology, and analytical chemistry.

Evolution of Luciferase

I and my co-workers has been shown that the luciferase in fireflies (Lampyridae, Coleoptera) was originated from “fatty acyl-CoA synthetase”, an enzyme involving in beta-oxidation (3-5). Recently, we demonstrated that changes in a few amino acids of fatty acyl-CoA synthetase in non-luminous beetle give significant luminescence activity (6). This result indicates that special catalytic activity of new enzyme is readily evolvable from unrelated common enzyme by small number of substitution steps.

Origin of Luciferin

With regard to the origin of luciferins, we showed that luciferin in sea-firefly (Cypridina hilgendorfii) is biosynthesized from three amino acids (Typ, Arg, Ile) (7,8), coelenterazine (luciferins and luminescence chromophore of photoproteins in various marine luminous organisms, such as jellyfishes and crustaceans) is biosynthesized from 2 Tyr and 1 Phe in the deep-sea copepod, Metridia pacifica (9), and firefly luciferin from 1,4-hydroquinone and 2 Cys in the Japanese firefly Luciola lateralis (10, 11). Luciferins are very specialized substrate for luminescence. But these our results show that some luciferins are biosynthesized from usual materials such as alpha-amino acids.

Evolution of Luminous Organisms

Molecular phylogenetic analyses of luminous beetles have also been performed. The result suggests that luminescence in the some species of click beetle (Elateridae) evolved once in the subfamily Agrypninae and common ancestor of Elateridae was probably non-luminous (12). In contrast, the family Lampyridae is the monophyletic group and the common ancestor of Lampyridae was luminous (13).

Gene Duplication in Luminous Organisms

We found that fireflies possess two different genes of luciferase. One is responsible for the yellow-luminescence of lantern in larvae and adult, on the other hand, another is responsible for the dim green-luminescence of egg and whole body of pupae. Phylogenetic analysis revealed that these two luciferase genes were originated at the basal position of the family Lampyridae by gene duplication (14-17).

Bioluminescence of Luminous Mushroom

We determined with Russian Academy of Science that fungal pre-luciferin is ‘hispidin’ (6-(3,4-dihydroxystyryl)-4-hydroxy-2-pyrone). We showed that luminous mushrooms emit continuous green light by the enzymatic reaction of 3-hydroxy hispidin (luciferin) and undetermined luciferase (18). We also showed the oxyluciferin structure and proposed the novel bioluminescence mechanism of luminous fungi with Russian Academy of Science and San Paulo University (19). Now we proposed that continuous glow of luminous mushroom is regulated by the slow biosynthetic recycling of hispidin from caffeic acid, a luminescence reaction product (20).

DNA Barcoding of Luminous Organisms

Now, I join in the Japanese DNA barcode of life project (http://www.jboli.org/en/). My tasks are the construction of barcoding database on Japanese Elateridae (21) and Japanese luminous animals (22).

 

REFERENCES

  1. Oba Y, Branham MA, Fukatsu T (2011) The terrestrial bioluminescent animals of Japan. Zool. Sci. 28: 771-789. (Review)
  2. Oba, Y. (2014) Insect Bioluminescence in the Post-Molecular Biology Era. pp. 88-114. In: Hoffman, K. H. (Ed.) Insect Molecular Biology and Ecology. CRC Press, Florida.
  3. Oba Y, Ojika M, Inouye S (2003) Firefly luciferase is a bifunctional enzyme: ATP-dependent monooxygenase and a long chain fatty acyl-CoA synthetase. FEBS Lett.540: 251-254.
  4. Oba Y, Sato M, Ohta Y, Inouye S (2006) Identification of paralogous genes of firefly luciferase in the Japanese firefly, Luciola cruciata. Gene368: 53-60.
  5. Oba Y (2009) On the origin of beetle luminescence. In “Bioluminescence in Focus- A Collection of Illuminating Essays” Ed by VB Meyer-Rochow, Research Signpost, Kerala, India, pp 277-290.
  6. Oba Y, Iida K, Inouye S (2009) Functional conversion of fatty acyl-CoA synthetase to firefly luciferase by site-directed mutagenesis: A key substitution responsible for luminescence activity. FEBS Lett.583: 2004-2008.
  7. Oba Y, Kato S, Ojika M, Inouye S (2002) Biosynthesis of luciferin in the sea firefly, Cypridina hilgendorfii: L-Tryptophan is a component in Cypridina luciferin. Tetrahedron Lett.43: 2389-2392.
  8. Kato S, Oba Y, Ojika M, Inouye S (2004) Identification of the biosynthetic units of Cypridina luciferin in Cypridina (Vargula) hilgendorfii by LC/ESI-TOF-MS. Tetrahedron60: 11427-11434.
  9. Oba Y, Kato S, Ojika M, Inouye S (2009) Biosynthesis of coelenterazine in the deep-sea copepod, Metridia pacifica. Biochem. Biophys. Res. Commun.390: 684-688.
  10. Oba Y, Yoshida N, Kanie S, Ojika M, Inouye S. (2013) Biosynthesis of firefly luciferin in adult lantern: decarboxylation of L-cysteine is a key step for benzothiazole ring formation in firefly luciferin. PLOS ONE 8: e84023.
  11. Kanie S, Nishikawa T, Ojika M, Oba Y (2016) One-pot non-enzymatic formation of firefly luciferin in a neutral buffer from p-benzoquinone and cysteine. Sci. Rept. 6: 24794.
  12. Sagegami-Oba R, Oba Y, Ohira H (2007) Phylogenetic relationships of click beetles (Coleoptera: Elateridae) inferred from 28S ribosomal DNA: Insights into the evolution of bioluminescence in Elateridae. Mol. Phylogenet. Evol.42: 410-421. 
  13. Sagegami-Oba R, Takahashi N, Oba Y (2007) The evolutionary process of bioluminescence and aposematism in cantharoid beetles (Coleoptera: Elateroidea) inferred by the analysis of 18S ribosomal DNA. Gene400: 104-113.
  14. Oba Y, Mori N, Yoshida M, Inouye S. (2010) Identification and characterization of a luciferase isotype in the Japanese firefly, Luciola cruciata, involving in the dim glow of firefly eggs. Biochemistry 49: 10788-10795.
  15. Oba Y, Furuhashi M, Bessho M, Sagawa S, Ikeya H, Inouye S. (2013) Bioluminescence of a firefly pupa: involvement of a luciferase isotype in the dim glow of pupae and eggs in the Japanese firefly, Luciola lateralis. Photochem. Photobiol. Sci. 12: 854-863.
  16. Bessho-Uehara M, Oba Y (2017) Identification and characterization of the Luc2-type luciferase in the Japanese firefly, Luciola parvula, involved in a dim luminescence in immobile stages. Luminescence (in press).
  17. Bessho-Uehara M, Konishi K, Oba Y (2017) Biochemical characteristics and gene expression profiles of two paralogous luciferases from the Japanese firefly Pyrocoelia atripennis (Coleoptera, Lampyridae, Lampyrinae): Insight into the evolution of firefly luciferase genes. Photochem. Photobiol. Sci. 16, 1301-1310.
  18. Purtov KV, Petushkov VN, Baranov MS, Mineev KS, Rodionova NS, Kaskova ZM, Tsarkova AS, Petunin AI, Bondar VS, Rodicheva EK, Medvedeva SE, Oba Y, Oba Y, Arseniev AS, Lukyanov S, Gitelson JI, Yampolsky IV (2015) The chemical basis of fungal bioluminescence. Angew. Chem. 54: 8124-8128.
  19. Kaskova ZM, Dörr, FA, Petushkov VN, Purtov KV, Tsarkova AS, Rodionova NS, Mineev KS, Guglya EB, Kotlobay A, Baleeva NS, Baranov MS, Arseniev AS, Gitelson JI, Lukyanov S, Suzuki Y, Kanie S, Pinto E, Mascio PD, Waldenmaier HE, Pereira TA, Carvalho RP, Oliveira AG, Oba Y, Bastos EL, Stevani CV, Yampolsky IV. (2017) Mechanism and color modulation of fungal bioluminescence. Sci. Adv. 3: e1602847.
  20. Oba Y, Suzuki Y, Martins GNR, Carvalho RP, Pereira TA, Waldenmaier HE, Kanie S, Naito M, Oliveira AG, Dörr FA, Pinto E, Yampolsky IV and Stevani CV (2017) Identification of hispidin as a bioluminescent active compound and its recycling biosynthesis in the lumimous fungal fruiting body. Photochem. Photobiol. Sci. (in press).
  21. Oba Y, Ohira H, Murase Y, Moriyama A, Kumazawa Y (2015) DNA barcoding of Japanese click beetles (Coleoptera, Elateridae). PLOS ONE 10: e0116612.
  22. Oba Y, Schultz TD (2014) Eco-Evo Bioluminescence on Land and in the Sea. pp. 3-36. In: Thouand, G and Marks, R. (Eds.). Bioluminescence: Fundamentals and Applications in Biotechnology Vol. 1. Springer, Heidelberg. (Review)

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