Zhu Xu-Dong, Shi Xiang, Wang Shu-Wen, Chu Ju, Zhu Wei-Hong, Ye Bang-Ce, Zuo Peng, Wang Yong-Hong
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology Shanghai 200237 China
Institute of Fine Chemicals, East China University of Science and Technology Shanghai 200237 China.
RSC Adv. 2019 Feb 5;9(8):4507-4513. doi: 10.1039/c8ra09684h. eCollection 2019 Jan 30.
A high-throughput screening system based on droplet microfluidic sorting was developed and employed for screening of high lactic acid-producing . In this system, water-in-oil-in-water (W/O/W) droplets, which were ∼12 pL in volume were used as picoliter-reactors for lactic acid fermentation. A fluorescent sensor was developed and used for monitoring pH which indicated the production of lactic acid. After fermentation, fluorescence activated cell sorting was performed with high sensitivity and speed. Using this microfluidic high-throughput screening system, we found a mutant with a yield of 76 g L lactic acid which was 52% higher than its parent strain with a screening throughput exceeding 10 clones per h.
开发了一种基于液滴微流控分选的高通量筛选系统，并用于筛选高产乳酸的菌株。在该系统中，体积约为12皮升的水包油包水（W/O/W）液滴被用作皮升规模的乳酸发酵反应器。开发了一种荧光传感器用于监测pH值，以此指示乳酸的产生。发酵后，以高灵敏度和速度进行荧光激活细胞分选。使用这种微流控高通量筛选系统，我们发现了一个突变体，其乳酸产量为76克/升，比其亲本菌株高出52%，筛选通量超过每小时10个克隆。
