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A selection platform for carbon chain elongation using the CoA-dependent pathway to produce linear higher alcohols

Paper ID Volume ID Publish Year Pages File Format Full-Text
31564 44819 2012 8 PDF Available
Title
A selection platform for carbon chain elongation using the CoA-dependent pathway to produce linear higher alcohols
Abstract

Production of green chemicals and fuels using metabolically engineered organisms has been a promising alternative to petroleum-based production. Higher chain alcohols (C4–C8) are of interest because they can be used as chemical feedstock as well as fuels. Recently, the feasibility of n-hexanol synthesis using Escherichia coli has been demonstrated by extending the modified Clostridium CoA-dependent n-butanol synthesis pathway, thereby elongating carbon chain length via reactions in reversed β-oxidation, (or β-reduction). Here, we developed an anaerobic growth selection platform that allows selection or enrichment of enzymes for increased synthesis of C6 and C8 linear alcohols. Using this selection, we were able to improve the carbon flux towards the synthesis of C6 and C8 acyl-CoA intermediates. Replacement of the original enzyme Clostridium acetobutylicum Hbd with Ralstonia eutropha homologue PaaH1 increased production of n-hexanol by 10-fold. Further directed evolution by random mutagenesis of PaaH1 improved n-hexanol and n-octanol production. This anaerobic growth selection platform may be useful for selecting enzymes for production of long-chain alcohols and acids using this CoA-dependent pathway.

► A selection platform was developed to favor chain elongation in linear alcohol synthesis. ► Ralstonia eutropha homolog PaaH1 increased production of n-hexanol by 10-fold. ► Directed evolution of PaaH1 using this selection improved n-hexanol and n-octanol production.

Keywords
n-hexanol; n-octanol; Biofuel; Protein directed evolution; Hexanoic acid; 3-hydroxyacyl-CoA dehydrogenase
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A selection platform for carbon chain elongation using the CoA-dependent pathway to produce linear higher alcohols
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 14, Issue 5, September 2012, Pages 504–511
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