模块化多基合成酶中的密集甲基表酶活性的解析,使用并列模块表酶试验
Xinqiang Xie1, Ashish Garg1, Chaitan Khosla2
1Department of Chemistry, Brown University , Box H, Providence, Rhode Island 02912-9108, United States.
Journal of the American Chemical Society
|July 7, 2017
概括
多基合成酶 (PKS) 中的脱水酶 (DH) 域可能具有隐藏的表皮酶活性. 研究人员在特定的DH域中发现了内在的表皮酶和脱水酶功能,挑战了以前关于它们作用的假设.
科学领域:
- 生物化学
- 分子生物学
- 酵素学
背景情况:
- 脱水酶 (DH) 域是多基合成酶 (PKS) 的组成部分.
- 这些域通常与在多基生物合成中产生表皮化中间体有关.
- 一些DH域的精确酶功能仍然是神秘的.
研究的目的:
- 在多基合成酶系统中研究特定脱水酶域的内在酶活性.
- 来自南菌素和尼日里辛PKS的DH域的表皮酶和脱水酶功能.
- 确定已知脱水酶活性的DH域是否也具有表皮酶功能.
主要方法:
- 使用双重平衡同位素交换 (EIX) 试验.
- 使用一个新开发的Tandem模块Epimerase测定.
- 分析了南胺素 (NanDH1,NanDH5,NanDH2) 和尼格里辛 (NigDH1) PKS,以及红色胺素合成酶 (EryDH4) 的DH域.
主要成果:
- NanDH1,NanDH5和NigDH1表现出内在的表皮酶活性.
- 这三个表皮酶活性DH域也意外地具有内在脱水酶活性.
- 传统的DH域 (EryDH4,NanDH2) 显示出神秘的表皮酶活性.
结论:
- 脱水酶域可以具有双重,内在的表皮酶和脱水酶活动.
- 在PKS中DH域的功能作用比以前理解的要多样化.
- 这一发现对理解多基生物合成和设计新型化合物有意义.
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