生物合成将非原生酸构建块纳入类产品,使用工程化基域
Fumihiro Ishikawa1, Maya Nohara1, Akimasa Miyanaga2
1Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-oskaa, Osaka 577-8502, Japan.
ACS chemical biology
|December 2, 2024
概括
工程酶通过接受新型酸来扩大非核糖体 (NRPs) 的多样性. 这项工作通过途径工程推进了通过途径工程构建定制的含酸代谢产物.
科学领域:
- 生物化学和分子生物学
- 代谢工程是代谢工程.
- 自然产品的合成自然产品的合成
背景情况:
- 非核糖体 (NRPs) 是具有治疗潜力的多样化的二次代谢物,由非核糖体合成酶 (NRPSs) 生合成.
- 酸是NRP结构多样性的关键组成部分.
- 之前的研究设计了EntE腺化域以接受单替代的酸.
研究的目的:
- 为了研究先前设计的EntE突变体 (N235G) 与各种非替代的酸衍生物的基质范围.
- 阐明突变EntE酶对基质识别的结构基础.
- 为了证明工程NRPS酶在生产新型含酸中的实用性.
主要方法:
- 在EntE腺化域的局部定向突变发生 (在235位的Asn-to-Gly替代).
- 使用各种替代的酸衍生物和非水解的阿里尔-AMP类似物进行酶分析.
- 对突变EntE (N235G) 结构分析与基质类似物复合.
- 工程EnTE突变体与肠杆菌素和活菌素生物合成酶的结合.
主要成果:
- 突变的EntE (N235G) 成功地适应了各种非替代的酸 (,甲基,甲氧,,) 和一个3-基替代的酸.
- 结构研究表明,突变EntE的扩大的基质结合口袋容纳了重和功能化的酸.
- 工程NRPS途径产生含有3-基酸,酸和3--2-基酸的.
结论:
- 改造的EntE (N235G) 突变体对酸激活具有扩展的基质特异性.
- 基质结合口袋的扩大对于容纳多样化的酸结构至关重要.
- NRPS路径工程为产生具有潜在治疗应用的新型酸含有代谢物提供了一个强大的策略.
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