Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides
Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides
复制标题
鉴定和消除醌修饰途径中增强球形红杆菌辅酶 Q(10) 产量的代谢瓶颈
DOI：
10.1016/j.ymben.2015.03.012
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发表时间：
2015-05-01
影响因子：
8.4
通讯作者：
Yu, Hongwei
中科院分区：
文献类型：
--
作者：
Lu, Wenqiang;Ye, Lidan;Yu, Hongwei
在本报告中，除了UbiG之外，还鉴定了醌修饰途径（QMP）中的UbiE和UbiH作为类球红细菌合成辅酶Q（10）的瓶颈酶。CoQ（10）含量在UbiE和UbiG共过表达后增加，然而，伴随着中间体10 P-MMBQ的积累。然后共过表达UbiH以将代谢通量拉向下游，导致CoQ（10）生产率升高和生物量降低。另一方面，调节UbiE和UbiG的表达水平以消除中间积累，然而以牺牲生产率为代价。为了减轻对生产力或细胞生长的不利影响，我们将UbiG与UbiE融合，并将它们定位在膜上以提高中间体转化。通过将UbiE和UbiG融合到pufX上，辅酶Q（10）的积累达到108.51 ± 2.76 rog/L，生物量为12.2 ± 0.9 g/L。因此，我们将优化的QMP和先前工程化的2-甲基-D-β-4-磷脂途径（MEP）组合以进一步促进CoQ（10）生物合成，从而产生具有138 +/-2.64 mg/L CoQ（10）产量的菌株。(C)2015年国际代谢工程学会。由爱思唯尔公司出版All rights reserved.
In this report, UbiE and UbiH in the quinone modification pathway (QMP) were identified in addition to UbiG as bottleneck enzymes in the CoQ(10) biosynthesis by Rhodobacter sphaeroides. The CoQ(10) content was enhanced after co-overexpression of UbiE and UbiG, however, accompanied by the accumulation of the intermediate 10P-MMBQ. UbiH was then co-overexpressed to pull the metabolic flux towards downstream, resulting in an elevated CoQ(10) productivity and decreased biomass. On the other hand, the expression levels of UbiE and UbiG were tuned to eliminate the intermediate accumulation, however at the sacrifice of productivity. To alleviate the detrimental effect on either productivity or cell growth, we fried to fuse UbiG with UbiE and localize them onto the membrane to elevate intermediate conversion. By fusing UbiE and UbiG to pufX, CoQ(10) was accumulated to 108.51 2.76 rog/L with a biomass of 12.2 +/- 0.9 g/L. AL Iasi, we combined the optimized QMP and the previously engineered 2-meillyl-D-erythritol-4-phosphaLe pathway (MEP) to further boost CoQ(10) biosynthesis, resulting in a strain with 138 +/- 2.64 mg/L CoQ(10) production. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.