摘要
Shikimic acid(SA) is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for expressing the plurality of the genes in one plasmid, so it is widely applied to increase the yield of metabolites. In the present study, on the basis of a shikimate kinase genetic defect strain Escherichia coli BL21(?aro L/aro K, DE3), the key enzyme genes aro G, aro B, tkt A and aro E of SA pathway were co-expressed and compared systematically by constructing a series of multi-gene expression vectors. The results showed that different gene co-expression combinations(two, three or four genes) or gene orders had different effects on the production of SA. SA production of the recombinant BL21-GBAE reached to 886.38 mg·L^(-1), which was 17-fold(P < 0.05) of the parent strain BL21(?aro L/aro K, DE3).
Shikimic acid （SA） is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for expressing the plurality of the genes in one plasmid, so it is widely applied to increase the yield of metabolites. In the present study, on the basis ofa shikimate kinase genetic defect strain Escherichia eoli BL21 （AaroL/aroK, DE3）, the key enzyme genes aroG, aroB, tktA and aroE of SA pathway were co-expressed and compared systematically by constructing a series of multi-gene expression vectors. The results showed that different gene co-expression combinations （two, three or four genes） or gene orders had different effects on the production of SA. SA production of the recombinant BL21-GBAE reached to 886.38 mg.L-1, which was 17-fold （P〈 0.05） of the parent strain BL21 （AaroL/aroK, DE3）.
基金
supported by National Important Special Foundation of the New Drug Development(2012ZX 09301002-003 and 2014ZX09201001-012)
Shanghai Innovation Action Plan of Science and Technology(14431905900)
