Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing, China.
Key Laboratory of Industrial Biocatalysis, Ministry of Education, Tsinghua University, Beijing, China.
Biotechnol Bioeng. 2020 Jun;117(6):1724-1737. doi: 10.1002/bit.27327. Epub 2020 Apr 9.
Conventional microbial cell cultivation techniques are typically labor intensive, low throughput, and poorlyparallelized, rendering them inefficient. The development of automated, modular microbial cell micro-cultivation systems, particularly those employing droplet microfluidics, have gained attention for their high-throughput, highly paralellized and efficient cultivation capabilities. Here, we report the development of a microbial microdroplet culture system (MMC), which is an integrated platform for automated, high-throughput cultivation and adaptive evolution of microorganisms. We demonstrated that the MMC yielded both accurate and reproducible results for the manipulation and detection of droplets. The superior performance of MMC for microbial cell cultivation was validated by comparing the growth curves of six microbial strains grown in MMC, conventional shake flasks or well plates. The highest incipient growth rate for all six microbial strains was achieved by using MMC. We also conducted an 18-day process of adaptive evolution of methanol-essential Escherichia coli strain in MMC and obtained two strains exhibiting higher growth rates compared with the parent strain. Our study demonstrates the power of MMC to provide an efficient and reliable approach for automated, high-throughput microbial cultivation and adaptive evolution.
传统的微生物细胞培养技术通常劳动强度大、通量低且难以平行化，效率低下。自动化、模块化的微生物细胞微培养系统的发展，特别是采用液滴微流控技术的系统，因其高通量、高度平行化和高效培养能力而受到关注。在这里，我们报告了一种微生物微滴培养系统 (MMC) 的开发，该系统是一个用于微生物自动化、高通量培养和适应性进化的集成平台。我们证明了 MMC 可以准确、可重复地对液滴进行操作和检测。通过比较在 MMC、传统摇瓶或孔板中生长的六种微生物菌株的生长曲线，验证了 MMC 对微生物细胞培养的优异性能。对于所有六种微生物菌株，MMC 都实现了最高的初始生长速率。我们还在 MMC 中进行了甲醇必需型大肠杆菌菌株 18 天的适应性进化过程，获得了两个与亲本菌株相比生长速度更高的菌株。我们的研究证明了 MMC 提供了一种高效可靠的自动化、高通量微生物培养和适应性进化方法的能力。
