title: Tibetan Firefly Luciferase with Low Temperature Adaptation.
abstract: Fireflies are widespread all over the world and a numerous numbers of luciferases have been isolated and characterized. In this study, we identified and characterized the luciferase and luciferase-like genes from a Tibetan firefly collected in Shangri-La, China. The altitude of this area is more than 3300 m. We saw this Tibetan firefly flying with strong luminescence after sunset at ~10°C. We analyzed the transcriptome of Tibetan firefly using head, thorax, abdomen (without light organ), and light organ tissue by RNA sequencing. We identified one luciferase gene, which was almost identical to luciferase from fireflies Pyrocoelia species, and expressed specifically in the light organ. Interestingly, the optimal temperature of the Tibetan firefly recombinant luciferase was 10°C. The Kmfor D-luciferin and ATP of the recombinant luciferase was 23 and 154 μm, respectively. The optimal pH was around 7.0-7.5. The emission peak was 556 nm at pH 8.0, while it shifted to 606 nm at pH 6.0. We also found a luciferase-like gene with 43% identical amino acids to the Tibetan firefly luciferase, which was scarcely expressed in any portion of the adult body. No luciferase activity was detected for this luciferase-like protein.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Mitani
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Futahashi
authors.affiliation: Kunming Key Laboratory of Hydroecology Restoration of Dianchi Lake, Key Laboratory of Special Biological Resource Development and Utilization of Universities in Yunnan Province, Department of Biological Science and Technology, Kunming University, Kunming, China.
authors.forename: Zichao
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Honorary Curator of Zoological Museum, 86th High School, Guangzhou, China.
authors.forename: Xingcai
authors.initials: X
authors.lastname: Liang
authors.affiliation: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.|DAILAB, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmiya
mesh_terms: D000222:Adaptation, Physiological; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002681:China; D003080:Cold Temperature; D048888:Fireflies / Q000145:classification / Q000201:enzymology / Q000502:physiology*; D049454:Firefly Luciferin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D049409:Luciferases, Firefly / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D049408:Luminescent Agents / Q000378:metabolism; D008297:Male; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA; D017386:Sequence Homology, Amino Acid; D018609:Tibet; D059467:Transcriptome
chemical_list: D049408:Luminescent Agents; D011994:Recombinant Proteins; D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly