Han Guoqiang, Xu Ning, Sun Xieping, Chen Jinzhao, Chen Chun, Wang Qing
Life Science and Technology Institute, Yangtze Normal University, Chongqing 408100, P. R. China.
School of Advanced Agriculture and Bioengineering, Yangtze Normal University, Chongqing 408100, P. R. China.
ACS Omega. 2020 Mar 6;5(10):4751-4758. doi: 10.1021/acsomega.9b02747. eCollection 2020 Mar 17.
As one of the branched-chain amino acids, l-valine is an essential nutrient for most mammalian species. In this study, the l-valine producer was first constructed. Additionally, an improved biosensor based on the Lrp-type transcriptional regulator and temperature-sensitive replication was built. Then, the strain was mutagenized by atmospheric and room temperature plasma. A sequential three-step procedure was carried out to screen l-valine-producing strains, including the fluorescence-activated cell sorting (FACS), 96-well plate screening, and flask fermentation. The final mutant HL2-7 obtained by screening produced 3.20 g/L of l-valine, which was 21.47% higher than the titer produced by the starting strain. This study demonstrates that the l-valine-producing mutants can be successfully isolated based on the Lrp sensor system in combination with FACS screening after random mutagenesis.
作为支链氨基酸之一,L-缬氨酸是大多数哺乳动物物种的必需营养素。在本研究中,首先构建了L-缬氨酸生产菌株。此外,构建了一种基于Lrp型转录调节因子和温度敏感复制的改进型生物传感器。然后,通过常压室温等离子体对该菌株进行诱变。采用连续三步程序筛选L-缬氨酸生产菌株,包括荧光激活细胞分选(FACS)、96孔板筛选和摇瓶发酵。通过筛选获得的最终突变体HL2-7产生了3.20 g/L的L-缬氨酸,比出发菌株的产量高21.47%。本研究表明,在随机诱变后,基于Lrp传感器系统结合FACS筛选可以成功分离出L-缬氨酸生产突变体。