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Title: Construction of a Robust Non-Oxidative Glycolysis in Model Organisms for n-Butanol Production

Abstract

The Embden-Meyerhoff-Parnas (EMP) pathway, commonly known as glycolysis, represents the fundamental biochemical infrastructure for sugar catabolism in almost all organisms, as it provides key components for biosynthesis, energy metabolism, and global regulation. EMP-based metabolism synthesizes three-carbon (C3) metabolites prior to two-carbon (C2) metabolites, and must emit one CO2 in the synthesis of the C2 building block, acetyl-CoA, a precursor for many industrially important products. Using rational design, genome editing, and evolution, here we replaced the native glycolytic pathways in Escherichia coli with the previously designed non-oxidative glycolysis (NOG), which bypasses initial C3 formation and directly generates stoichiometric amounts of C2 metabolites. The resulting strain, which contains 11 gene overexpressions, 10 gene deletions by design, and more than 50 genomic mutations (including 3 global regulators) through evolution, grows aerobically in glucose minimal medium, but can ferment anaerobically to products with nearly complete carbon conservation. We confirmed that the strain metabolizes glucose through NOG by 13C tracer experiments. This re-designed E. coli strain represents a different approach for carbon catabolism, and may serve as a useful platform for bioproduction.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2]
  1. Univ. of California, Los Angeles, CA (United States). Dept. of Chemical and Biomolecular Engineering
  2. Univ. of California, Los Angeles, CA (United States). Dept. of Chemical and Biomolecular Engineering; Academia Sinica, Taipei (Taiwan)
Publication Date:
Research Org.:
Univ. of California, Los Angeles, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23). Biological Systems Science Division
OSTI Identifier:
1506427
Report Number(s):
DOE-UCLA-12384-1
DOE Contract Number:  
SC0012384; SC0006698
Resource Type:
Technical Report
Resource Relation:
Related Information: https://doi.org/10.1073/pnas.1802191115
Country of Publication:
United States
Language:
English
Subject:
E. coli; glycolysis; metabolic engineering; synthetic biology

Citation Formats

Lin, Paul P., Jaeger, Alec J., Wu, Tung-Yun, Xu, Sharon C., Lee, Abraxa S., Gao, Fanke, Chen, Po-Wei, and Liao, James C. Construction of a Robust Non-Oxidative Glycolysis in Model Organisms for n-Butanol Production. United States: N. p., 2019. Web. doi:10.2172/1506427.
Lin, Paul P., Jaeger, Alec J., Wu, Tung-Yun, Xu, Sharon C., Lee, Abraxa S., Gao, Fanke, Chen, Po-Wei, & Liao, James C. Construction of a Robust Non-Oxidative Glycolysis in Model Organisms for n-Butanol Production. United States. doi:10.2172/1506427.
Lin, Paul P., Jaeger, Alec J., Wu, Tung-Yun, Xu, Sharon C., Lee, Abraxa S., Gao, Fanke, Chen, Po-Wei, and Liao, James C. Wed . "Construction of a Robust Non-Oxidative Glycolysis in Model Organisms for n-Butanol Production". United States. doi:10.2172/1506427. https://www.osti.gov/servlets/purl/1506427.
@article{osti_1506427,
title = {Construction of a Robust Non-Oxidative Glycolysis in Model Organisms for n-Butanol Production},
author = {Lin, Paul P. and Jaeger, Alec J. and Wu, Tung-Yun and Xu, Sharon C. and Lee, Abraxa S. and Gao, Fanke and Chen, Po-Wei and Liao, James C.},
abstractNote = {The Embden-Meyerhoff-Parnas (EMP) pathway, commonly known as glycolysis, represents the fundamental biochemical infrastructure for sugar catabolism in almost all organisms, as it provides key components for biosynthesis, energy metabolism, and global regulation. EMP-based metabolism synthesizes three-carbon (C3) metabolites prior to two-carbon (C2) metabolites, and must emit one CO2 in the synthesis of the C2 building block, acetyl-CoA, a precursor for many industrially important products. Using rational design, genome editing, and evolution, here we replaced the native glycolytic pathways in Escherichia coli with the previously designed non-oxidative glycolysis (NOG), which bypasses initial C3 formation and directly generates stoichiometric amounts of C2 metabolites. The resulting strain, which contains 11 gene overexpressions, 10 gene deletions by design, and more than 50 genomic mutations (including 3 global regulators) through evolution, grows aerobically in glucose minimal medium, but can ferment anaerobically to products with nearly complete carbon conservation. We confirmed that the strain metabolizes glucose through NOG by 13C tracer experiments. This re-designed E. coli strain represents a different approach for carbon catabolism, and may serve as a useful platform for bioproduction.},
doi = {10.2172/1506427},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {4}
}