The Effect of E. coli Uridine-Cytidine Kinase Gene Deletion on Cytidine Synthesis and Transcriptome Analysis
The Effect of E. coli Uridine-Cytidine Kinase Gene Deletion on Cytidine Synthesis and Transcriptome Analysis
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大肠杆菌尿苷-胞苷激酶基因缺失对胞苷合成和转录组分析的影响
DOI：
10.3390/fermentation8110586
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发表时间：
2022-10
期刊：
影响因子：
--
通讯作者：
Haitian Fang
中科院分区：
文献类型：
--
作者：
Fengmin Liu;Tong Ye;Xiangjun Zhang;Cong Ma;Huiyan Liu;Haitian Fang
胞苷是一种抗病毒和抗癌药物中间体，其主要生产方法是发酵。尿苷-胞苷激酶（UCK）催化胞苷磷酸化生成胞苷酸的逆过程，影响大肠杆菌胞苷生物合成途径中胞苷的积累。胞苷产生菌E.以大肠杆菌NXBG-11为出发菌株，采用CRISPR/Cas9技术从染色体基因组中敲除编码UCK的udk基因。突变株E. coliNXBG-12，并对其转录组学进行研究，以了解udk基因缺失对胞苷合成和全细胞转录的影响。突变株E. coli NXBG-12胞苷产量是出发菌株E. coliNXBG-11在37 °C下摇瓶发酵40 h后，敲除udk基因，转录组分析显示，突变株与原始株差异表达基因共1168个，上调基因559个，下调基因609个。初步表明，UDK基因敲除对胞苷合成网络有积极影响，因为参与胞苷合成的基因显著上调（P
Cytidine is an antiviral and anticancer drug intermediate, its primary method of manufacture being fermentation. Uridine-cytidine kinase (UCK) catalyzes the reverse process of phosphorylation of cytidine to produce cytidylic acid, which influences cytidine accumulation in the Escherichia coli cytidine biosynthesis pathway. The cytidine-producing strain E. coli NXBG-11 was used as the starting strain in this work; the udk gene coding UCK was knocked out of the chromosomal genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology. The mutant strain E. coli NXBG-12 was obtained; its transcriptomics were studied to see how udk gene deletion affected cytidine synthesis and cell-wide transcription. The mutant strain E. coli NXBG-12 generated 1.28 times more cytidine than the original strain E. coli NXBG-11 after 40 h of shake-flask fermentation at 37 °C. The udk gene was knocked out, and transcriptome analysis showed that there were 1168 differentially expressed genes between the mutant and original strains, 559 upregulated genes and 609 downregulated genes. It was primarily shown that udk gene knockout has a positive impact on the cytidine synthesis network because genes involved in cytidine synthesis were significantly upregulated (p
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