State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
Biotechnol Appl Biochem. 2020 May;67(3):414-429. doi: 10.1002/bab.1890. Epub 2020 Jan 31.
Wild-type Escherichia coli usually does not accumulate l-threonine, but E. coli strain TWF001 could produce 30.35 g/L l-threonine after 23-H fed-batch fermentation. To understand the mechanism for the high yield of l-threonine production in TWF001, transcriptomic analyses of the TWF001 cell samples collected at the logarithmic and stationary phases were performed, using the wild-type E. coli strain W3110 as the control. Compared with W3110, 1739 and 2361 genes were differentially transcribed in the logarithmic and stationary phases, respectively. Most genes related to the biosynthesis of l-threonine were significantly upregulated. Some key genes related to the NAD(P)H regeneration were upregulated. Many genes relevant to glycolysis and TCA cycle were downregulated. The key genes involved in the l-threonine degradation were downregulated. The gene rhtA encoding the l-threonine exporter was upregulated, whereas the genes sstT and tdcC encoding the l-threonine importer were downregulated. The upregulated genes in the glutamate pathway might form an amino-providing loop, which is beneficial for the high yield of l-threonine production. Many genes encoding the 30S and 50S subunits of ribosomes were also upregulated. The findings are useful for gene engineering to increase l-threonine production in E. coli.
野生型大肠杆菌通常不会积累 l-苏氨酸,但大肠杆菌 TWF001 菌株在 23-H 流加发酵后可生产 30.35 g/L 的 l-苏氨酸。为了了解 TWF001 生产 l-苏氨酸高产的机制,我们对对数期和静止期的 TWF001 细胞样本进行了转录组分析,以野生型大肠杆菌 W3110 菌株作为对照。与 W3110 相比,对数期和静止期分别有 1739 个和 2361 个基因差异转录。大多数与 l-苏氨酸生物合成相关的基因均显著上调。一些与 NAD(P)H 再生相关的关键基因上调。许多与糖酵解和 TCA 循环相关的基因下调。与 l-苏氨酸降解相关的关键基因下调。编码 l-苏氨酸外排泵的 rhtA 基因上调,而编码 l-苏氨酸输入蛋白的 sstT 和 tdcC 基因下调。谷氨酸途径中上调的基因可能形成一个提供氨基酸的循环,这有利于 l-苏氨酸的高产。许多编码 30S 和 50S 核糖体亚基的基因也上调。这些发现有助于大肠杆菌中 l-苏氨酸生产的基因工程。