Enhancement of cytidine production by coexpression of gnd, zwf, and prs genes in recombinant Escherichia coli CYT15
Enhancement of cytidine production by coexpression of gnd, zwf, and prs genes in recombinant Escherichia coli CYT15
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DOI：
10.1007/s10529-012-1068-3
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发表时间：
2013-02-01
影响因子：
2.7
通讯作者：
Chen, Ning
中科院分区：
文献类型：
--
作者：
Fang, Haitian;Xie, Xixian;Chen, Ning
胞苷是几种抗病毒药物的前体。戊糖磷酸途径（PPP）主要负责NADPH和5-磷酸- α -d-核糖1-二磷酸，是大肠杆菌胞苷生物合成的重要前体。为了提高胞苷的产量，我们获得了重组大肠杆菌CYT15-gnd-prs-zwf，该基因共表达了分别编码磷酸核糖基焦磷酸合成酶、葡萄糖-6-磷酸脱氢酶和6-磷酸葡萄糖酸脱氢酶（PPP的三个关键酶）的prs、zwf和gnd基因。在发酵实验中，菌株CYT15-gnd-prs-zwf以葡萄糖为底物，产胞苷735 mg /l，约为。比亲本菌株CYT15的胞苷产量高128%。zwf、gnd和prs的共表达略微降低了生长（3.2%），增加了葡萄糖摄取（72%）。这是首次报道大肠杆菌通过PPP增加代谢通量从而增加胞苷产量的研究。
Cytidine is a precursor of several antiviral drugs. The pentose phosphate pathway (PPP) is primarily responsible for NADPH and 5-phospho-alpha-d-ribose 1-diphosphate as an important precursor of cytidine biosynthesis in Escherichia coli. To enhance cytidine production, we obtained the recombinant E. coli CYT15-gnd-prs-zwf that co-expressed the prs, zwf, and gnd genes encoding phosphoribosylpyrophosphate synthetase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase (three key enzymes in PPP) respectively. In fermentation experiments, strain CYT15-gnd-prs-zwf produced 735 mg cytidine/l using glucose as substrate, which was approx. 128 % higher than the cytidine production by the parental strain (CYT15). Co-expression of zwf, gnd, and prs decreased growth (3.2 %) slightly and increased glucose uptake (72 %). This is the first study to report increased cytidine production by increasing metabolic flux through the PPP in E. coli.