通过同源重组进行有效的模块化多基合成酶的演变
Joseph A Chemler, Ashootosh Tripathi, Douglas A Hansen
1Sequoia Sciences, Inc. , 1912 Innerbelt Business Center Drive, Saint Louis, Missouri 63114, United States.
Journal of the American Chemical Society
|August 1, 2015
概括
研究人员设计了混合多基合成酶 (PKS) 来制造新的宏和宏. 这种模块化方法可以通过组装PKS路径来生产定制的抗生素样分子.
科学领域:
- 生物化学
- 合成生物学
- 自然产品合成
背景情况:
- I型多基合成酶 (PKS) 是大型的多功能酶,负责生产复杂的天然产品.
- PKS的模块化结构允许通过修改酶域来设计定制生物催化剂.
- 之前的PKS工程工作往往导致酶活性降低和产品产量受限.
研究的目的:
- 开发一种用于生成和识别功能性化学PKS酶的多功能方法.
- 使用工程 PKS 合成定制的宏和宏.
- 探索PKS工程创造新型抗生素样分子的潜力.
主要方法:
- 来自Saccharomyces cerevisiae中的皮克罗米辛和红色素通路的PKS基因混合,以创建混合库.
- 采用96孔板式的高通量等离子体净化,转化,测序,蛋白质表达,体外反应和代谢物分析.
- 产生和选的仿制PKS酶,以获得新的功能和代谢物产生.
主要成果:
- 成功生成并鉴定出具有新功能的活体PKS酶.
- 表达这些模拟PKS的Streptomyces venezuelae菌株产生了工程化巨乳素.
- 通过进一步功能化由选定的PKS仿真体产生的巨乳素,合成了一种新型的巨乳素仿真体.
结论:
- 开发的方法为设计PKS路径作为模块化组件提供了一个强大的平台.
- 这种方法有助于创建具有潜在治疗应用的定制宏和宏.
- PKS的模块化组装为发现和生产新的类似抗生素的化合物提供了有前途的策略.
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