title: Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides.
abstract: In this report, UbiE and UbiH in the quinone modification pathway (QMP) were identified in addition to UbiG as bottleneck enzymes in the CoQ10 biosynthesis by Rhodobacter sphaeroides. The CoQ10 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 CoQ10 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 tried to fuse UbiG with UbiE and localize them onto the membrane to elevate intermediate conversion. By fusing UbiE and UbiG to pufX, CoQ10 was accumulated to 108.51±2.76mg/L with a biomass of 12.2±0.9g/L. At last, we combined the optimized QMP and the previously engineered 2-methyl-d-erythritol-4-phosphate pathway (MEP) to further boost CoQ10 biosynthesis, resulting in a strain with 138±2.64mg/L CoQ10 production.
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Wenqiang
authors: W
authors: Lu
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University, Hangzhou 310027, China.
authors: Lidan
authors: L
authors: Ye
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Xiaomei
authors: X
authors: Lv
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Wenping
authors: W
authors: Xie
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Jiali
authors: J
authors: Gu
authors: Group of Bioengineering, ZheJiang NHU Company Limited, Shaoxing 312521, China.
authors: Zhaofeng
authors: Z
authors: Chen
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Yongqiang
authors: Y
authors: Zhu
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors: Aipeng
authors: A
authors: Li
authors: Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China. Electronic address: yuhongwei@zju.edu.cn.
authors: Hongwei
authors: H
authors: Yu
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D008780:Methyltransferases / Q000096:biosynthesis / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000096:biosynthesis / Q000235:genetics; D007652:Oxo-Acid-Lyases / Q000096:biosynthesis / Q000235:genetics; D012242:Rhodobacter sphaeroides / Q000235:genetics / Q000378:metabolism; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D014451:Ubiquinone; D006899:Mixed Function Oxygenases; D008780:Methyltransferases; D007652:Oxo-Acid-Lyases; C075960:chorismate pyruvate lyase; C024989:coenzyme Q10
keywords: Bottleneck removal; CoQ(10); Fusion expression; Intermediate elimination; Quinone modification pathway; Rhodobacter sphaeroides