一种对androst-4-ene-3,17-dione反应的非自然转录因子的动态设计
Ming Zhao1, Mengkai Hu1, Rumeng Han1
1Anhui Engineering Laboratory for Industrial Microbiology Molecular Breeding, College of Biology and Food Engineering, Anhui Polytechnic University, Wuhu 241000, China.
Synthetic and systems biotechnology
|April 15, 2024
概括
研究人员设计了一种人工转录因子 (TF),该转录因子对androst-4-ene-3,17-dione (AD) 产生反应. 这种新型TF增强了微生物细胞工厂的AD生产,克服了自然TF的局限性.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 蛋白质工程是指蛋白质工程.
背景情况:
- 微生物细胞工厂需要转录因子 (TFs) 在类固醇生产中进行代谢调节.
- 现有的微生物系统缺乏有效的TF,这些TF对androst-4-ene-3,17-dione (AD) 特别作出反应.
- 识别对AD有反应的自然TF是具有挑战性和资源密集的.
研究的目的:
- 设计和制造一种新型的人工转录因子 (TF),该因子能够特别结合并对AD作出反应.
- 通过人工TF设计一种微生物系统,以增强AD的产生.
- 展示一种可推广的策略,用于制造非自然TF元素用于代谢工程.
主要方法:
- 结构引导分子动力学 (MD) 模拟用于设计人工TF (AdT).
- 作为一个分子开关,设计了一种AD-响应性联体结合域 (LBD).
- 该ADT组装了DNA结合和转录激活域,并在基于酵母的AD生物传感器中进行了测试.
主要成果:
- 基于AD结合的MD模拟,成功设计了一种人工TF,AdT.
- 一个使用AdT的酵母生物传感器证明了其在响应AD时的功能.
- 一种变体,AdT F320Y,显示转录因子活性增加了1.44倍.
- 该研究为AD微生物细胞工厂创造了新的TF元素.
结论:
- 开发的人工TF (AdT) 为AD代谢工程提供了一个新的工具.
- 设计策略为微生物细胞工厂创建定制TF提供了一种方法.
- 这种方法预计将适用于宿主细胞中的其他信号通路.
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