Genetic engineering of reveromycin A biosynthetic gene cluster and creation of its derivatives

瑞维霉素A生物合成基因簇的基因工程及其衍生物的创建

• 批准号: 18580085
• 负责人: TAKAHASHI Shunji
• 金额: $ 2.57万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580085/
• 关键词: Reveromycin A; Polyketide; P450

Reveromycin A, which is a polyketide compound produced by Streptomyces reveromyceticus, inhibits bone resorption by inducing apoptosis specifically in osteoclasts. To obtain new bioactive derivatives of reveromycin A, the cloning of reveromycin A biosynthetic gene cluster was conducted. Based on the information from mRNA expression pattern and genome shotgun sequencing data, the polyketide biosynthetic gene cluster responsible for reveromycin A biosynthesis was obtained from S. reveromyceticus genome. The sequence analysis revealed the additional polyketide starter module that incorporates methylmalonyl CoA. This suggests that reveromycin A is biosynthesized with additional propionate unit. Interestingly, the reveromycin derivative (reveromycin A2) supporting the DNA sequence information was isolated from the secondary metabolites from S. reveromyceticus. In the biosynthetic gene cluster, P450 which catalyzes the C-18 hydroxylation of reveromycin biosynthetic intermediate was identified. Because the enzyme is a target protein to create reveromycin A derivatives, the crystal graphic analysis is now in progress. The function of unknown genes in the cluster is also investigated through biochemical analysis and gene knock out.

炎霉素A是由链霉菌产生的聚酮化合物化合物，通过特异性诱导破骨细胞的凋亡来抑制骨吸收。为了获得炎霉素A的新生物活性衍生物，进行了霉素A的克隆A生物合成基因簇。基于来自mRNA表达模式和基因组shot弹枪测序数据的信息，负责炎霉素的聚酮化合物生物合成基因簇从牛rove菌基因组中获得了生物合成。序列分析揭示了融合了甲基丙二酰基COA的附加聚酮启动器模块。这表明炎霉素A具有额外的丙酸单元生物合成。有趣的是，从牛rove菌的次生代谢产物中分离出支持DNA序列信息的炎霉素衍生物（Reverymycin A2）。在生物合成基因簇中，鉴定了促霉素生物合成中间体的C-18羟基化的P450。由于该酶是产生丙霉素A衍生物的靶蛋白，因此现在正在进行晶体图形分析。还通过生化分析和基因淘汰，还研究了簇中未知基因的功能。

项目成果

蛋白質核酸酵素 ケミカルバイオロジー

蛋白质核酸酶化学生物学

• 发表时间: 2007
• 作者: 高橋 俊二;Shunji Takahashi;高橋 俊二;Shunji Takahashi;加藤 直樹
• 通讯作者: 加藤 直樹

蛋白質核酸酵素ケミカルバイオロジー

蛋白质核酸酶化学生物学

• 发表时间: 2007
• 作者: 高橋 俊二;Shunji Takahashi;高橋 俊二;Shunji Takahashi;加藤 直樹;加藤 直樹
• 通讯作者: 加藤 直樹

リベロマイシンA生合成遺伝子クラスターの解析

利博霉素A生物合成基因簇分析

• 发表时间: 2007
• 作者: 高橋 俊二;他
• 通讯作者: 他