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Modular optimization of multi-gene pathways for fumarate production

Paper ID Volume ID Publish Year Pages File Format Full-Text
31478 44800 2016 10 PDF Available
Title
Modular optimization of multi-gene pathways for fumarate production
Abstract

•Modular pathway engineering was used to produce fumarate.•Fusion proteins were constructed to improve the production of fumarate.•Gene expression strengths cassettes were optimized to increase fumarate.•DNA-guided scaffolds and RNA switchs were used to enhance fumarate production.•The final strain TGFA091-16 could produce 33.13 g/L fumarate.

Microbial fumarate production from renewable feedstock is a promising and sustainable alternative to petroleum-based chemical synthesis. Here, we report a modular engineering approach that systematically removed metabolic pathway bottlenecks and led to significant titer improvements in a multi-gene fumarate metabolic pathway. On the basis of central pathway architecture, yeast fumarate biosynthesis was re-cast into three modules: reduction module, oxidation module, and byproduct module. We targeted reduction module and oxidation module to the cytoplasm and the mitochondria, respectively. Combinatorially tuning pathway efficiency by constructing protein fusions RoMDH-P160A and KGD2-SUCLG2 and optimizing metabolic balance by controlling genes RoPYC, RoMDH-P160A, KGD2-SUCLG2 and SDH1 expression strengths led to significantly improved fumarate production (20.46 g/L). In byproduct module, synthetizing DNA-guided scaffolds and designing sRNA switchs enabled further production improvement up to 33.13 g/L. These results suggest that modular pathway engineering can systematically optimize biosynthesis pathways to enable an efficient production of fumarate.

Keywords
Fumarate; Modular pathway engineering; Multi-gene pathway; Synthetic biology
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 33, January 2016, Pages 76–85
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