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Title: Engineering glucose metabolism of Escherichia coli under nitrogen starvation

A major aspect of microbial metabolic engineering is the development of chassis hosts that have favorable global metabolic phenotypes, and can be further engineered to produce a variety of compounds. In this work, we focus on the problem of decoupling growth and production in the model bacterium Escherichia coli, and in particular on the maintenance of active metabolism during nitrogen-limited stationary phase. We find that by overexpressing the enzyme PtsI, a component of the glucose uptake system that is inhibited by α-ketoglutarate during nitrogen limitation, we are able to achieve a fourfold increase in metabolic rates. Alternative systems were also tested: chimeric PtsI proteins hypothesized to be insensitive to α-ketoglutarate did not improve metabolic rates under the conditions tested, whereas systems based on the galactose permease GalP suffered from energy stress and extreme sensitivity to expression level. Overexpression of PtsI is likely to be a useful arrow in the metabolic engineer’s quiver as productivity of engineered pathways becomes limited by central metabolic rates during stationary phase production processes.
Authors:
 [1] ; ORCiD logo [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [2] ;  [1]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Univ. of California, Berkeley, CA (United States). Dept. of Chemical & Biomolecular Engineering, Dept. of Bioengineering; Technical Univ. of Denmark, Hørsholm (Denmark). Novo Nordisk Foundation Center for Biosustainability
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
npj Systems Biology and Applications
Additional Journal Information:
Journal Volume: 3; Journal Issue: 1; Journal ID: ISSN 2056-7189
Publisher:
Springer Nature
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1474995

Chubukov, Victor, Desmarais, John James, Wang, George, Chan, Leanne Jade G., Baidoo, Edward EK, Petzold, Christopher J., Keasling, Jay D., and Mukhopadhyay, Aindrila. Engineering glucose metabolism of Escherichia coli under nitrogen starvation. United States: N. p., Web. doi:10.1038/npjsba.2016.35.
Chubukov, Victor, Desmarais, John James, Wang, George, Chan, Leanne Jade G., Baidoo, Edward EK, Petzold, Christopher J., Keasling, Jay D., & Mukhopadhyay, Aindrila. Engineering glucose metabolism of Escherichia coli under nitrogen starvation. United States. doi:10.1038/npjsba.2016.35.
Chubukov, Victor, Desmarais, John James, Wang, George, Chan, Leanne Jade G., Baidoo, Edward EK, Petzold, Christopher J., Keasling, Jay D., and Mukhopadhyay, Aindrila. 2017. "Engineering glucose metabolism of Escherichia coli under nitrogen starvation". United States. doi:10.1038/npjsba.2016.35. https://www.osti.gov/servlets/purl/1474995.
@article{osti_1474995,
title = {Engineering glucose metabolism of Escherichia coli under nitrogen starvation},
author = {Chubukov, Victor and Desmarais, John James and Wang, George and Chan, Leanne Jade G. and Baidoo, Edward EK and Petzold, Christopher J. and Keasling, Jay D. and Mukhopadhyay, Aindrila},
abstractNote = {A major aspect of microbial metabolic engineering is the development of chassis hosts that have favorable global metabolic phenotypes, and can be further engineered to produce a variety of compounds. In this work, we focus on the problem of decoupling growth and production in the model bacterium Escherichia coli, and in particular on the maintenance of active metabolism during nitrogen-limited stationary phase. We find that by overexpressing the enzyme PtsI, a component of the glucose uptake system that is inhibited by α-ketoglutarate during nitrogen limitation, we are able to achieve a fourfold increase in metabolic rates. Alternative systems were also tested: chimeric PtsI proteins hypothesized to be insensitive to α-ketoglutarate did not improve metabolic rates under the conditions tested, whereas systems based on the galactose permease GalP suffered from energy stress and extreme sensitivity to expression level. Overexpression of PtsI is likely to be a useful arrow in the metabolic engineer’s quiver as productivity of engineered pathways becomes limited by central metabolic rates during stationary phase production processes.},
doi = {10.1038/npjsba.2016.35},
journal = {npj Systems Biology and Applications},
number = 1,
volume = 3,
place = {United States},
year = {2017},
month = {1}
}

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