在库拉生物合成中通过多基基酸脱水域的维尼洛基脱水
William D Fiers1, Greg J Dodge2, David H Sherman3
1Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Journal of the American Chemical Society
|December 15, 2016
概括
聚胺合成脱水酶 (DH) 域是产生新型化合物的关键. 这项研究揭示了古拉途径DH的新机制和高活性,为合成生物学应用提供了潜力.
科学领域:
- 生物化学
- 合成生物学
- 酵素学
背景情况:
- 聚基合成酶 (PKS) 对新疗法和化学物质至关重要.
- 在PKS中脱水酶 (DH) 域对于多基基的烯酸形成至关重要.
- 对DH机制的有限理解阻碍了PKS对合成生物学进行重新设计.
研究的目的:
- 调查库拉通路中的DH域的机制和动力学.
- 描述PKS DH酶的基质特异性和选择性.
- 为 PKS 工程提供对烯形成的结构见解.
主要方法:
- 在实验室中使用合成基质对四个库拉辛通路DH域进行表征.
- 运动分析和LC-MS/MS用于活动测量.
- 一个CurJ-DH基质复合物的X射线晶体学.
主要成果:
- 对于全跨三酸盐的形成,CurK-DH具有很高的催化效率 (kcat为72s-1).
- 对CurJ和CurH DHs提出了一种新型立体特异性机制,其中涉及一种乙烯基酸中介物.
- 结构数据显示了对氨酸形成至关重要的酶基质相互作用.
结论:
- 库拉辛途径DHs为理解多基基脱水提供了一个模型.
- 新的催化机制和高活性为合成生物学提供了机会.
- 了解DH对于合理设计PKS的各种应用至关重要.
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