Investigation of Unutilized Bioluminescent Systems and Fluorescent Proteins, and Development of New Molecular Proves.

未利用的生物发光系统和荧光蛋白的研究以及新分子证据的开发。

• 批准号: 15404009
• 负责人: NIWA Haruki
• 金额: $ 8.45万
• 依托单位: The University of Electro-Communications
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15404009/
• 关键词: bioluminescence; luminous worm; Odotosyllis; luminous snail; Latia; liciferin; liciferase; 分離; 構造決定; 蛍光性; 抽出; 発光生物; 精製

1)From the marine luminescent worm, Odontosyll is phosphorea collected off from Mission Bay, San Diego, U.S.A., the light producing substance, Odontosyll is luciferin was isolated under the guidance of luciferin-luciferase reaction. The bioluminescence spectrum (λmax 500nm) using the isolated luciferin and the crude luciferase was similar to that of live specimens. The UV spectrum was essentially identical with that reported previously by Shimomura. The molecular formula of the luciferin was determined from the high resolution ESI mass spectrum to be C_<15>H_8O_<12>S_3.2)Exploration of luminescent marine organisms was performed at New Caledonia, some luminous worm and sea hare was found.3)From the fresh-water limpet like snail, Latia neritoides collected at the Nugtunu stream near Mt. Pirongia, New Zealand, the luciferase was isolated under the guidance of luciferin-luciferase reaction. The detailed biochemical studies indicated that the active form of the luciferase was found to be a hexamer of N-linked glycoprotein monomer having the molecular mass 31kDa. The cloning of the luciferase gene was succeeded. However heterologous expression of the gene was not succeeded so far.

1)从美国圣地亚哥观澜湾采集的海洋发光虫Odontosyll is荧光素中，通过荧光素-荧光素酶反应分离得到发光物质Odontosyll is Luciferin。用分离的荧光素和粗制的荧光素酶的生物发光光谱(λmax 500 nm)与活体标本相似。紫外光谱与Shimomura之前报道的结果基本相同。经高分辨电喷雾质谱测定，其分子式为C_&lt；15&gt；H_8O_&lt；12&gt；S_3)对新喀里多尼亚的发光海洋生物进行了考察，发现了一些发光的蠕虫和海兔。3)从努格图努山附近的努格图努溪采集的象蜗牛的淡水帽贝中发现了Latia neritodes。Pirongia，新西兰，在荧光素-荧光素酶反应的指导下分离得到荧光素酶。详细的生化研究表明，荧光素酶的活性形式是N-连接的糖蛋白单体的六角体，分子量为31 kDa。成功克隆了荧光素酶基因。然而，到目前为止，该基因的异源表达尚未成功。

项目成果

天然型L-システインまたはその誘導体を用いたホタル発光基質の生合成システム及び本システムを含んだ発光基質溶液

使用天然L-半胱氨酸或其衍生物的萤火虫发光底物生物合成系统以及含有该系统的发光底物溶液

• 发表时间: 2004

Chemiluminescence f 6-aryl-2 methylimidazo[1,2-a]pyrazin-3(7H)-ones in DMSO/TMG and in diglyme/acetate buffer : support for the chemiexcitation process to generate the singlet-excited state of neutral oxyluciferin in high quantum yield in the Cypridina(Va

DMSO/TMG 和二甘醇二甲醚/乙酸盐缓冲液中 6-芳基-2 甲基咪唑[1,2-a]吡嗪-3(7H)-酮的化学发光：支持化学激发过程，以在中性氧化荧光素中产生单线态激发态

• 发表时间: 2006
• 期刊: Tetrahedron Lett. Vol.47
• 作者: 平野 誉;Mitsuhiro Nakamuraら;Kotaro Mori ら;Yuko Takahashi ら
• 通讯作者: Yuko Takahashi ら

Real light emitter in the bioluminescence of the calcium-activated photoproteins aequorin and obelin: light emission from the singlet-excited state of coelenteramide phenolate anion in a contact ion pair

• DOI: 10.1016/j.tet.2006.04.044
• 发表时间: 2006-06-26
• 期刊: TETRAHEDRON
• 影响因子: 2.1
• 作者: Mori, Kotaro;Maki, Shojiro;Hirano, Takashi
• 通讯作者: Hirano, Takashi

Bioluminescence Activity of Latia Luciferin Analogues : Replacement of the 2,6,6-Trimethylcyclohexene Ring into the Methyl-substituted Phenyl Groups.

Latia 荧光素类似物的生物发光活性：将 2,6,6-三甲基环己烯环替换为甲基取代的苯基。

• 发表时间: 2005
• 期刊: Tetrahedron Lett. 46, No.1
• 作者: Mitsuhiro Nakamura;Masashi Mamino;Mizuki Masaki;Shojiro Maki;Ryo Matsui;Satoshi Kojima;Takashi Hirano;Yoshihiro Ohmiya;Haruki Niwa.
• 通讯作者: Haruki Niwa.

6,8-Diarylimidazo[11,2a]pyrazin-3(7H)-ones as Potential Chemiluminescent pH/Superoxide Double Sensor.

6,8-二芳基咪唑并[11,2a]吡嗪-3(7H)-酮作为潜在的化学发光 pH/超氧化物双传感器。

• 发表时间: 2005
• 期刊: Bioluminescence and Chemiluminescence(ed by A.Tsuji, M.Matsumoto, M.Maeda, L.J.Kricka, P.E.Stanley)(World Scientific), Singapore
• 作者: R.Saito;N.Suga;A.Katoh;T.Hirano;H.Niwa.
• 通讯作者: H.Niwa.