クロロフィル分解の分子機構

叶绿素分解的分子机制

• 批准号: 00J07856
• 负责人: 鈴木 康予
• 金额: $ 1.73万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 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/grant/KAKENHI-PROJECT-00J07856/
• 关键词: クロロフィル分解; ピロフェオホルビドの生成; フェオホルビダーゼ; ダイコン

一部の植物では,クロロフィル分解の初期反応においてフェオホルビドaはピロフェオホルビドaへと分解されることが知られている.この反応は維管束植物では酵素的な加水分解と自発的な脱炭酸反応の2段階の反応による.この加水分解反応を触媒する酵素をフェオホルビダーゼと名付け,研究を行っている.本酵素は老化によって誘導されるものと構成的に存在するものの2種類があり,FPLCを用いて高度に精製をした.ダイコン(Raphanus sativus)の子葉から,老化誘導型,構成型はそれぞれ10,000倍と6,500倍に精製できた.フェオホルビダーゼのサブユニットは77kDaであるが,サンプル調製時に熱処理することで,SDS-PAGEにおいて16.8,15.9,11.8kDaの3つのバンドを得た.高度な精製標品の断片化されたペプチドを用いて部分アミノ酸配列の解析を行い,2種類のアミノ酸配列の決定に成功した.決定したアミノ酸配列をもとにプライマーを設計し,RT-PCRによって部分ヌクレオチド配列を得た.さらに,3'-RACE,5'-RACEを組み合わせて,フェオホルビダーゼ遺伝子のクローニングを行っている.現在までに明らかになったヌクレオチド配列を用いてBLASTサーチを行ったところ,ダイコンのフェオホルビダーゼと高い相同性を持つものがシロイヌナズナに存在し,このことは酵素活性の植物分布とも一致していた.今後は,大腸菌を用いた大量発現系の構築を行い,得られたタンパクを用いて抗体作成を行う.また,ウエスタン,ノーザン解析を行い,フェオホルビダーゼの発現調節機構を解明していく予定である.

A part of the plant is divided into two parts: the first part is divided into three parts: the first part is divided into four parts: the first part is divided into three parts: the first part is divided into four parts: the first part is divided into three parts: the second part is divided into four parts: the first part is divided into three parts: the first part is divided into four parts: the first part is divided into four parts: the second part is divided into four parts: the first part is divided into four parts: the first part is divided into four parts: the second part is divided into four parts: the first part is divided into four parts: the first part is divided into four parts: the second part is divided into four parts: the first part is divided into four parts: the second part is divided into four parts: the third part is divided The two stages of decarboxylation are the two stages of decarboxylation. The enzyme of this hydrolysis reaction was studied in detail. The enzyme is highly refined in the presence of two types of enzymes. The cotyledons of Raphanus sativus are senescence induced, and the constitutive type is refined by 10,000 times and 6,500 times. The protein is 77kDa and the protein is 16.8,15.9,11.8kDa in SDS-PAGE. The analysis of highly refined standard fragments was carried out successfully in the determination of partial acid alignment. To determine the sequence of DNA sequences,RT-PCR was used. Today, the 3'-RACE,5'-RACE group is composed of two groups, one group is composed of two groups, the other group is composed of three groups, and the other group is composed of three groups. Now, the enzyme activity of the plant is consistent with the distribution of the plant. In the future, the construction of a large number of detection systems for Escherichia coli will be carried out, and the production of antibodies will be carried out. For example, if you want to analyze the situation, you can analyze the situation and adjust the mechanism.

项目成果

Suzuki, Y., Doi, M., Shioi, Y.: "Two enzymatic reaction pathways in the formation of pyropheophorbide a"Photosynthesis Research. 74. 225-233 (2002)

Suzuki, Y.、Doi, M.、Shioi, Y.：“焦脱镁叶绿酸 a 形成中的两种酶反应途径”光合作用研究。