通过依赖于酸盐的酶多样化林科萨胺生物合成的分子基础
Takahiro Mori1,2,3,4, Yoshitaka Moriwaki5,6,7, Kosuke Sakurada8
1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan. tmori@mol.f.u-tokyo.ac.jp.
依赖于皮里多-5'-酸盐 (PLP) 的酶LmbF和CcbF使林科萨米德的抗生素途径多样化. 结构分析显示,活性部位的芳香残留物决定了不同的催化机制,使酶功能切换和创建新衍生物成为可能.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 有机化学 有机化学
背景情况:
- 林科萨米德抗生素林科米辛A和切莱斯提的生物合成涉及PLP依赖的酶LmbF和CcbF.
- 这些酶通过催化类似基质上的不同反应来分离生物合成途径.
研究的目的:
- 阐明LmbF和CcbF不同的催化机制的结构基础.
- 调查活性现场残留物在基质识别和反应结果中的作用.
- 为了设计这些酶的选择性和氧气利用.
主要方法:
- 在X射线晶体学.
- 酶对接模拟的模拟
- 分子动力学模拟的模拟.
- 局部导向的突变发生.
主要成果:
- 活动部位的芳香残留被确定为基质结合和反应结果的关键决定因素.
- 突变性研究成功地将CcbF的催化功能转换为LmbF的催化功能.
- 对LmbF和CcbF的工程变体表现出新的氧化-化活性,产生非自然的林科萨胺衍生物.
结论:
- 结构洞察力提供了对在林科萨胺生物合成中的PLP依赖酶多样化机制的理解.
- 对LmbF和CcbF的合理工程使得能够创建具有改变催化活性和新产品形成的酶.
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