区域选择性控制SAM化通过结构引导的氧化基转移酶的定向进化
Yixun Jiang1,2,3, Jianqiang Feng4, Haoran Niu1,2
1Frontiers Science Center for Synthetic Biology, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
研究人员将氧化腺转移酶 (HATase) 转化为区域选择性酶. 这一突破可以精确控制S-adenosylmethionine (SAM) 裂变,扩大化学合成的酶工具.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 合成生物学 合成生物学
- 生物化学 生物化学
背景情况:
- 酶化提供了温和的反应条件,与传统的化学方法形成鲜明对比.
- 在抗生素生物合成中,S-adenosylmethionine (SAM) 是SAM依赖的类酶的关键辅因子.
- 氧化腺转移酶 (HATases) 和依赖SAM的类酶共享DUF-62蛋白家族,但HATases使用的是水,而不是SAM.
研究的目的:
- 设计一个HATase用于区域选择性控制SAM化,这是以前没有取得的壮举.
- 识别关键蛋白质残留物和突变,这些突变决定了SAM裂解中的区域选择性.
- 探索DUF-62超级家族在区域选择性SAM修饰方面的潜力.
主要方法:
- 来自 *Thermotoga maritima* MSB8.8. 的 HATase 的蛋白质工程.
- 使用*大肠杆菌*细胞表面显示来产生功能突变的定向进化.
- 计算研究分析突变对化物导向和SAM裂变区域选择性的影响.
主要成果:
- 工程HATase突变物成功地将SAM转化为甲基化物 (CH3I) 和S-adenosylhomocysteine (SAH).
- 确定了影响SAM区域选择性分裂的关键残留物.
- 这些突变为其他HATases赋予了区域选择性化能力,证明了广泛的适用性.
结论:
- 这项研究证明了HATases的成功工程用于区域选择性SAM裂变,创造了新的化物甲基转移酶.
- 这项工作突出了DUF-62蛋白家族对于精确SAM修改的潜力.
- 工程酶扩大了生物催化和化学合成的工具包.
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