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Direct conversion of glucose to malate by synthetic metabolic engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
23558 43452 2013 7 PDF Available
Title
Direct conversion of glucose to malate by synthetic metabolic engineering
Abstract

Synthetic metabolic engineering enables us to construct an in vitro artificial synthetic pathways specialized for chemical manufacturing through the simple heat-treatment of the recombinant mesophiles having thermophilic enzymes, followed by rational combination of those biocatalytic modules. In this work, we constructed a synthetic pathway capable of direct conversion of glucose to malate. The reversible carboxylation of pyruvate catalyzed by a malic enzyme derived from Thermococcus kodakarensis (TkME) (ΔG°′ = +7.3 kJ mol−1) was coupled with a thermodynamically favorable non-ATP-forming Embden–Meyerhof pathway to balance the consumption and regeneration of redox cofactors and to shift the overall equilibrium toward malate production (glucose + 2HCO3− + 2H → 2 malate + 2H2O; ΔG°′ = −121.4 kJ mol−1). TkME exhibited both pyruvate carboxylation (malate-forming) and pyruvate reduction (lactate-forming) activities. By increasing HCO3− concentration, the reaction specificity could be redirected to malate production. As a result, the direct conversion of glucose to malate was achieved with a molar yield of 60%.

► An in vitro metabolic pathway for conversion of glucose to malate was constructed. ► Reversible carboxylation was coupled with a non-ATP-forming glycolysis pathway. ► Thermococcus malic enzyme showed both lactate-forming and malate-forming activities. ► Reaction specificity was redirected to malate by increasing the HCO3− concentration.

Keywords
Synthetic metabolic engineering; Thermophilic enzyme; Malic enzyme; Carbon dioxide fixation; Thermodynamic equilibrium
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 164, Issue 1, 10 March 2013, Pages 34–40
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