通过组合工程评估和利用更新的聚基酸合成酶模块
Katherine A Ray1, Joshua D Lutgens1, Ramesh Bista1
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, TX.
Research square
|August 7, 2023
概括
使用更新的模块边界对聚基酸合成酶的组合工程显示出有希望. 一个新的平台有效地构建了大型合成酶,揭示了挑战,并使新型化合物的混合设计成为可能.
科学领域:
- 合成生物学 合成生物学
- 生物化学 生物化学
- 代谢工程是代谢工程.
背景情况:
- 聚乙组装线是模块化的,对于生产有价值的化合物至关重要.
- 组合工程提供了一条通往新型多基菌的路线.
- 之前的研究集中在短合成酶上;使用更新模块边界的较大合成酶尚未被探索.
研究的目的:
- 开发一个平台,用于构建和测试使用更新的模块界限使用更大的多基基合成酶.
- 研究从皮克罗米合成酶中获得的模块的组合可能性.
- 确定局限性并使混合合成酶的创建成为可能.
主要方法:
- 设计和实施了一个类似于BioBricks的平台,用于快速组装聚基酸合成酶.
- 从Pikromycin合成酶模块中构建并测试了5个三基,25个四基和125个五基合成酶.
- 结合了来自Erythromycin,Spinosyn和Rapamycin合成酶的模块,以创建混合结构.
主要成果:
- 从60%的三化物,32%的四化物和6.4%的五化物结构中获得了功能合成.
- 合成酶守门和模块跳转被确定为主要障碍.
- 成功创建混合合成酶,包括那些具有放松的合成酶,从Rapamycin合成酶保持门.
结论:
- 开发的平台可以高效地构建和测试大型的组合工程聚基合成酶.
- 了解诸如合成酶守门等局限性是提高功能性产量的关键.
- 混合合成酶,特别是那些利用轻松守门的合成酶,为生产设计者多基虫提供了令人兴奋的可能性.
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