Significantly enhanced production of isoprene by ordered coexpression of genes dxs, dxr, and idi in Escherichia coli
Significantly enhanced production of isoprene by ordered coexpression of genes dxs, dxr, and idi in Escherichia coli
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通过在大肠杆菌中有序共表达 dxs、dxr 和 idi 基因，显着提高异戊二烯的产量
DOI：
10.1007/s00253-012-4485-2
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发表时间：
2013-03-01
影响因子：
5
通讯作者：
Yu, Hongwei
中科院分区：
文献类型：
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作者：
Lv, Xiaomei;Xu, Haoming;Yu, Hongwei
我们通过引入白杨的异戊二烯合酶（ispS），构建了大肠杆菌中异戊二烯生产的生物合成途径。过表达 1-脱氧-d-木酮糖 5-磷酸合酶 (dxs)、1-脱氧-d-木酮糖 5-磷酸还原异构酶 (dxr) 和异戊烯基二磷酸 (IPP) 异构酶 (idi) 以增强异戊二烯产量。与重组BL21（pET-30a-ispS）相比，异戊二烯产量分别提高了0.65、0.16和1.22倍，并且发现idi是异戊二烯产量的关键调节点。为了优化大肠杆菌中异戊二烯的生产，我们尝试基于 pET-30a 构建具有不同顺序的基因 dxs、dxr 和 idi 的多顺反子操纵子。用 pET-30a-dxs/dxr/idi 转化的大肠杆菌获得了最高的异戊二烯产量，为 2.727 mg g−1 h−1（每干重）。有趣的是，发现基因顺序与代谢途径一致。这表明基因的顺序是代谢工程中的一个重要问题，并且可以根据生物合成途径优化顺序表达模式以实现有效的产物合成。
We constructed a biosynthetic pathway of isoprene production in Escherichia coli by introducing isoprene synthase (ispS) from Populus alba. 1-deoxy-d-xylulose 5-phosphate synthase (dxs), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (dxr) and isopentenyl diphosphate (IPP) isomerase (idi) were overexpressed to enhance the isoprene production. The isoprene production was improved 0.65, 0.16, and 1.22 fold over the recombinant BL21 (pET-30a-ispS), respectively, and idi was found to be a key regulating point for isoprene production. In order to optimize the production of isoprene in E. coli, we attempted to construct polycistronic operons based on pET-30a with genes dxs, dxr, and idi in various orders. The highest isoprene production yield of 2.727 mg g−1 h−1 (per dry weight) was achieved by E. coli transformed with pET-30a-dxs/dxr/idi. Interestingly, the gene order was found to be consistent with that of the metabolic pathway. This indicates that order of genes is a significant concern in metabolic engineering and a sequential expression pattern can be optimized according to the biosynthetic pathway for efficient product synthesis.