Enhanced cytidine production by a recombinant Escherichia coli strain using genetic manipulation strategies
Enhanced cytidine production by a recombinant Escherichia coli strain using genetic manipulation strategies
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DOI：
10.1007/s13213-013-0760-4
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发表时间：
2014-09-01
影响因子：
3
通讯作者：
Chen, Ning
中科院分区：
文献类型：
--
作者：
Fang, Haitian;Zhang, Chenglin;Chen, Ning
胞苷是一种核苷分子，被广泛用作抗病毒药物的前体。本研究以大肠杆菌K-12为基础，采用三步遗传操作策略，培养了胞苷产菌Cyt18。首先从大肠杆菌K-12株中删除胞苷脱氨酶基因（cdd），形成Cyt10。此外，从Cyt10菌株中灭活同源丝氨酸脱氢酶基因（thrA），形成Cyt12，其胞内天冬氨酸浓度有望增加。构建含有解淀粉芽孢杆菌CYTI中pyrB-pyrA操纵子的重组质粒pMG1105，将其导入Cyt12中，获得Cyt18菌株。与Cyt12菌株相比，重组菌株Cyt18的胞苷产量增加了约3倍（722.9 mg/l比249.3 mg/l）。
Cytidine is a nucleoside molecule that is widely used as a precursor for antiviral drugs. In this study, a cytidine-producing strain Cyt18 was developed from Escherichia coli K-12 through 3-step genetic manipulation strategies. Cytidine deaminase gene (cdd) was firstly deleted from the E. coli K-12 strain to develop Cyt10. Furthermore, homoserine dehydrogenase gene (thrA) was inactivated from the Cyt10 strain to develop Cyt12, in which the intracellular aspartate concentration was expected to be increased. The recombinant plasmid pMG1105 containing an pyrB-pyrA operon from Bacillus amyloliquefaciens CYTI was constructed and was introduced into Cyt12 to obtain the Cyt18 strain. Compared to the Cyt12 strain, the cytidine production by the recombinant strain Cyt18 was increased by similar to 3-fold (722.9 mg/l vs. 249.3 mg/l).