Document Detail


Engineering Escherichia coli for production of C₁₂-C₁₄ polyhydroxyalkanoate from glucose.
MedLine Citation:
PMID:  23141473     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Demand for sustainable materials motivates the development of microorganisms capable of synthesizing products from renewable substrates. A challenge to commercial production of polyhydroxyalkanoates (PHA), microbially derived polyesters, is engineering metabolic pathways to produce a polymer with the desired monomer composition from an unrelated and renewable source. Here, we demonstrate a metabolic pathway for converting glucose into medium-chain-length (mcl)-PHA composed primarily of 3-hydroxydodecanoate monomers. This pathway combines fatty acid biosynthesis, an acyl-ACP thioesterase to generate desired C₁₂ and C₁₄ fatty acids, β-oxidation for conversion of fatty acids to (R)-3-hydroxyacyl-CoAs, and a PHA polymerase. A key finding is that Escherichia coli expresses multiple copies of enzymes involved in β-oxidation under aerobic conditions. To produce polyhydroxydodecanoate, an acyl-ACP thioesterase (BTE), an enoyl-CoA hydratase (phaJ3), and mcl-PHA polymerase (phaC2) were overexpressed in E. coli ΔfadRABIJ. Yields were improved through expression of an acyl-CoA synthetase resulting in production over 15% CDW--the highest reported production of mcl-PHA of a defined composition from an unrelated carbon source.
Authors:
Daniel E Agnew; Amanda K Stevermer; J Tyler Youngquist; Brian F Pfleger
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Metabolic engineering     Volume:  14     ISSN:  1096-7184     ISO Abbreviation:  Metab. Eng.     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-12     Completed Date:  2013-05-09     Revised Date:  2013-12-04    
Medline Journal Info:
Nlm Unique ID:  9815657     Medline TA:  Metab Eng     Country:  Belgium    
Other Details:
Languages:  eng     Pagination:  705-13     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Cloning, Molecular / methods
Escherichia coli / metabolism*
Glucose / metabolism*
Multienzyme Complexes / genetics,  metabolism*
Polyhydroxyalkanoates / genetics,  isolation & purification,  metabolism*
Protein Engineering / methods*
Pseudomonas aeruginosa / physiology*
Recombinant Proteins / metabolism
Grant Support
ID/Acronym/Agency:
5T32GM08349/GM/NIGMS NIH HHS; T32 GM008349/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Multienzyme Complexes; 0/Polyhydroxyalkanoates; 0/Recombinant Proteins; IY9XDZ35W2/Glucose
Comments/Corrections

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