天然色素アントシアニンの合成とその機能の解明

天然色素花青素的合成及其功能阐明

• 批准号: 12460052
• 负责人: KONDO Tadao
• 金额: $ 7.42万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460052/
• 关键词: Synthesis of anthocyanin; acylated anthocyanin; reduction with Zn-Hg; Flav-3-ene; light shielding effect; heavenly blue anthocyanin; thimidine dimmer; damage of gene; チミジン二重体; アントシアニン; 超分子; メタロアントシアニン; グルコシル化; 紫外線防御; チミジン2量体; プロトデルフィン; キラル

Synthesis of anthocyanin, a natural pigment, and clarification of its biological functionAnthocyanin is major flower pigment and might play role of amount biological functions such as gene protection form sunlight, entomophily, and radical scavenger. It is used for a food colorant and eye therapeutic medicine.We developed the new synthesis (1) of acylated anthocyanins from flavonol glycoside and proven protection (2) of DNA by acylated anthocyanin (HBA) from sunlight, UV-A and B.(1) We realized the direct transformation to anthocyanin from rutin and its derivatives, a flavonol glycoside by using metal reduction reaction. Various reaction conditions were applied to the reduction. Rutin was treated with Zn-Hg in absolute 3% HCl-methanol at 0℃ to give cyanidin 3-rutinoside in 24% yield. And also, perbenzoylrutins were reduced to afford the corresponding anthocyanin in Ox(3E)5% yield. Zn powder in replacing of Zn-Hg gave flav-3-ene 3-rutinoside in 65% yield. Therefore, we found the new efficient synthetic route to a natural flav-3-ene.(2) We provided chemical evidence that heavenly blue anthocyanin (HBA), a pigment of blue petal of Ipomoea tricolor, has a light shielding effect of UV-A and B to protect the DNA. That HBA exists at 10-2M in epidermis cell of the petal, was clarified by micro-photometery of a blue protoplast, which was isolated after treatment of the tissue with cellulase. We built up a model system of the petal for protection of DNA from UV-light exposure. The DNA damage was analyzed by ELISA method using antibody of thymidine dimers. The concentration (10-3M) of an aq. HBA solution just as that of living cell completely blocked the DNA damage under UV-irradiation, indicating that the anthocyanin pigment in open flower has a function of protection of the gene from sun-light.

天然色素花色苷的合成及其生物学功能的阐明花色苷是花卉的主要色素，具有基因保护、抗光、抗虫、清除自由基等多种生物学功能。我们开发了（1）从黄酮醇糖苷合成酰化花青素的新方法，并证明了（2）酰化花青素（HBA）对日光、UV-A和B的保护作用。(1)利用金属还原反应实现了芦丁及其衍生物-黄酮醇糖苷直接转化为花色苷。将各种反应条件应用于还原。芦丁在无水3%盐酸-甲醇中，于0℃下与锌汞化合物反应，得到矢车菊素3-芸香糖苷，产率24%。此外，过苯甲酰芸香苷被还原以得到相应的花色苷，Ox（3E）产率为5%。用锌粉代替锌汞，得到黄酮-3-烯-3-芸香糖苷，收率65%。因此，我们找到了一条新的高效合成天然黄-3-烯的路线。(2)从化学上证明了三色牵牛花蓝色花瓣色素天兰花青素（HBA）对UV-A和B具有一定的屏蔽作用，对DNA具有一定的保护作用。经纤维素酶处理后分离出的蓝色原生质体，经显微光度测定，证实HBA存在于花瓣表皮细胞中，浓度为10 ~（-2）M。我们建立了一个模式系统的花瓣保护DNA从紫外线照射。用胸苷二聚体抗体ELISA法检测DNA损伤。将aq. HBA溶液与活细胞溶液一样，完全阻断了紫外线对DNA的损伤，表明开放花中的花青素具有保护基因免受阳光照射的功能。

项目成果

Kumi Yoshida: "Structure of Muscarinin A, an Acylated Anthocyanin, from Purplish Blue Spicate Flower Petals of Muscari arumeniacum"ITE Letters. 3. 35-38 (2002)

Kumi Yoshida：“毒蕈碱 A 的结构，一种酰化花青素，来自 Muscari arumeniacum 的紫蓝色穗状花花瓣”ITE Letters。

Kumi Yoshida: "Prevention of Uv-light Induced E, Z-isomerization of Caffeoyl Residues in the Diacylated Anthocyanin, Gentiodelphin, by Intramolecular Stacking"Tetrahedron Letters. 43. 6181-6184 (2002)

Kumi Yoshida：“通过分子内堆积防止二酰化花青素、龙胆海豚中咖啡酰残基的紫外线诱导 E、Z 异构化”四面体字母。

Rikke Norbaek: "Flavone C-Glycoside, Phenolic Acid, and Nitrogen Contents in Leaves of Barley Subject to Organic Fertilization Treatments"Journal of Agricultural and Food Chemistry. 51. 809-813 (2003)

Rikke Norbaek：“有机肥处理大麦叶子中的黄酮 C-糖苷、酚酸和氮含量”农业与食品化学杂志。

Kumi Yoshida: "Influence of E, Z-isomerization and Stability of Acylated Anthocyanins under the UV Irradiation"Biochemical Engineering Journal. (印刷中). (2003)

Kumi Yoshida：“紫外线照射下酰化花青素的 E、Z 异构化和稳定性的影响”生物化学工程杂志（2003 年出版）。

Yuki Toyama-Kato: "Analysis of Metal Elements of Hydrangea Sepals at Various growing Stages by ICP-AES"Biochemical Engineering Journal. (印刷中). (2003)

Yuki Toyama-Kato：“通过 ICP-AES 分析绣球花萼片的金属元素”（正在出版）。