Wang Jian-feng, Meng Hai-lin, Xiong Zhi-qiang, Zhang Si-liang, Wang Yong
Biotechnol Lett. 2014 May;36(5):1021-7. doi: 10.1007/s10529-014-1460-2.
Discovery of novel potential genetic targets to increase the supply of isoprenoid precursors, isopentyl/dimethylallyl diphosphate, is of importance for microbial production of isoprenoids. Here, to improve isoprenoid precursor supply, a flux distribution comparison analysis, based on the genome-scale model, was utilized to simultaneously predict the knockout, down- and up-regulated targets in Escherichia coli. 51 targets were in silico discovered. All knockout and up-regulated targets were experimentally tested to enhance lycopene production. Five knockout targets (deoB, yhfw, yahI, pta and eutD) and four up-regulated targets (ompN, ompE, ndk and cmk) led to 10-45% increases of lycopene yield, respectively, which had not been uncovered in previous studies. When engineering of the five most significant targets gdhA, eutD, tpiA, ompE and ompN, were combined the lycopene titer improved by 174% in shake-flask and 81% in bioreactor fermentations with a maximum yield of 454 mg l(-1).
发现新型潜在基因靶点以增加类异戊二烯前体（异戊烯基/二甲基烯丙基二磷酸）的供应，对于微生物生产类异戊二烯具有重要意义。在此，为了改善类异戊二烯前体的供应，基于基因组规模模型进行了通量分布比较分析，以同时预测大肠杆菌中的基因敲除、下调和上调靶点。通过计算机模拟发现了51个靶点。对所有基因敲除和上调靶点进行了实验测试，以提高番茄红素的产量。五个基因敲除靶点（deoB、yhfw、yahI、pta和eutD）和四个上调靶点（ompN、ompE、ndk和cmk）分别使番茄红素产量提高了10 - 45%，这在以前的研究中尚未发现。当对五个最重要的靶点gdhA、eutD、tpiA、ompE和ompN进行工程改造时，摇瓶发酵中番茄红素产量提高了174%，生物反应器发酵中提高了81%，最高产量达到454 mg l(-1)。
