Liu Xianglei, Lin Jun, Hu Haifeng, Zhou Bin, Zhu Baoquan
State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai, 200040, China.
World J Microbiol Biotechnol. 2014 Sep;30(9):2543-50. doi: 10.1007/s11274-014-1679-z. Epub 2014 Jun 4.
Shikimic acid (SA) is the key synthetic material of Oseltamivir, which is an effective drug for the prevention and treatment of influenza. In this study, to block the downstream metabolic pathway of SA, the shikimate kinase isoenzyme genes aroK and aroL were deleted by Red recombination. Moreover, the key enzyme genes aroG, aroB, tktA and aroE of SA pathway were co-expressed by constructing the recombinant vector pETDuet-GBAE. As a result, SA production of E. coli BW25113 (∆aroL/aroK, DE3)/pETDuet-GBAE reached 1,077.6 mg/l when low amounts of sorbitol (5 g/l) were fed in shake flasks. The yield was 3.7 times that when glucose was used (P < 0.05). The results showed that sorbitol was an optimized carbon source for the high efficient accumulation of SA for the first time, which was applicable to use in the industry for high yields and low consumption.
莽草酸（SA）是奥司他韦的关键合成原料，奥司他韦是一种预防和治疗流感的有效药物。在本研究中，为阻断SA的下游代谢途径，通过Red重组缺失了莽草酸激酶同工酶基因aroK和aroL。此外，通过构建重组载体pETDuet-GBAE共表达了SA途径的关键酶基因aroG、aroB、tktA和aroE。结果，在摇瓶中补加少量山梨醇（5 g/l）时，大肠杆菌BW25113（∆aroL/aroK，DE3）/pETDuet-GBAE的SA产量达到1077.6 mg/l。该产量是使用葡萄糖时的3.7倍（P < 0.05）。结果首次表明山梨醇是SA高效积累的优化碳源，适用于工业上的高产低耗。
