质量激活滴滴分类用于选择生产素的大肠杆菌
Emory M Payne1, Bridget E Murray1, Laura I Penabad1
1Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48103, United States.
Analytical chemistry
|October 11, 2023
概括
合成生物学使用质量激活滴滴分类 (MADS) 来选工程细胞. 该方法利用电喷离子质谱法 (ESI-MS),使化学产生的微生物变体的高通量选择成为可能.
科学领域:
- 合成生物学 合成生物学
- 生物技术是生物技术.
- 分析化学 分析化学
背景情况:
- 工程细胞的理想性质,如化学生产,对于合成生物学至关重要.
- 选和分类变体库是细胞工程中常见的瓶.
- 滴滴微流体提供高通量分析,但传统上依赖于光检测.
研究的目的:
- 为了证明质量激活滴滴分类 (MADS) 的应用,用于工程细胞变体的高通量选.
- 为了评估MADS使用用于素生产的工程Escherichia大肠杆菌.
- 为了根据质谱学信号来选择和恢复特定的细胞变体.
主要方法:
- 使用滴滴微流体来封装工程大肠杆菌在20nL的滴滴中.
- 采用电子喷射电离质谱仪 (ESI-MS) 来检测液滴中细胞产生的氨酸.
- 实施了质量激活滴滴分类 (MADS) 以根据ESI-MS信号强度在0.5样本/秒时选择滴滴.
主要成果:
- 在微流体滴中证明了大肠杆菌的成功生长和氨酸生产.
- 表明大肠杆菌产生的氨酸可以通过ESI-MS在液滴中检测到.
- 在MADS选择后,高氨酸产生的变体的纯度达到了96-98%.
- 启用了对排序的细胞进行下游验证的恢复.
结论:
- 质量激活滴滴分类 (MADS) 是合成生物学中高通量选的有效方法.
- MADS克服了光分析在细胞工程液滴微流体学中的局限性.
- 该技术可用于全细胞工程和生物催化剂生产.
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