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Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides

Paper ID Volume ID Publish Year Pages File Format Full-Text
31527 44807 2015 9 PDF Available
Title
Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides
Abstract

•UbiE and UbiH were identified as bottleneck enzymes in the CoQ10 biosynthesis.•The quinone modification pathway (QMP) was finely tuned with different strategies.•Accumulation of intermediate revealed the blocking in the pathway.•Biomass and the productivity were the important parameters in metabolic engineering.•200% improvement in CoQ10 titer after co-engineering the MEP and QMP.

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±2.76 mg/L with a biomass of 12.2±0.9 g/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±2.64 mg/L CoQ10 production.

Keywords
Quinone modification pathway; CoQ10; Rhodobacter sphaeroides; Fusion expression; Bottleneck removal; Intermediate elimination
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Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 29, May 2015, Pages 208–216
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us