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Title: Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes

Abstract

This study constructed cell‐free glycolytic enzyme systems and compared them to their in vivo functions in Escherichia coli . Under in vitro conditions, flux regulation followed enzyme concentrations and kinetics. In E. coli, only one of the isozymes of phosphofructokinase (PfkA) and fructose‐bisphosphate aldolase (FbaA) facilitate Embden‐Meyerhof‐Parnas (EMP) flux, but under in vitro assays, these isozymes were interchangeable. Additionally, in vitro introduction of the Entner–Doudoroff (ED) pathway improved glycolysis rates, while in vivo overexpression of the ED pathway could not capture significant flux unless its phosphotransferase system (PTS) was knocked out. Lastly, in vivo dynamic 13 C‐experiments revealed that the labeling order of EMP pathway intermediates was not strictly cascade, indicating intracellular metabolites were not well mixed. These enigmatic observations cannot be fully explained by thermodynamics or substrate level regulations. This article supports the long‐time conjecture that EMP enzymes are channeled, and the PTS may be an anchor point to initiate enzyme assemblies. © 2018 American Institute of Chemical Engineers AIChE J , 65: 483–490, 2019

Authors:
 [1];  [2];  [1];  [3];  [3]; ORCiD logo [2]; ORCiD logo [1]
  1. Dept. of Energy, Environmental and Chemical Engineering Washington University St. Louis MO 63130
  2. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 China
  3. Lawrence Berkeley National Laboratory Emeryville CA 64608
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1472164
Grant/Contract Number:  
DESC0018324
Resource Type:
Publisher's Accepted Manuscript
Journal Name:
AIChE Journal
Additional Journal Information:
Journal Name: AIChE Journal Journal Volume: 65 Journal Issue: 2; Journal ID: ISSN 0001-1541
Publisher:
Wiley Blackwell (John Wiley & Sons)
Country of Publication:
United States
Language:
English

Citation Formats

Abernathy, Mary H., Zhang, Yuchen, Hollinshead, Whitney D., Wang, George, Baidoo, Edward E. K., Liu, Tiangang, and Tang, Yinjie J. Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes. United States: N. p., 2018. Web. doi:10.1002/aic.16367.
Abernathy, Mary H., Zhang, Yuchen, Hollinshead, Whitney D., Wang, George, Baidoo, Edward E. K., Liu, Tiangang, & Tang, Yinjie J. Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes. United States. https://doi.org/10.1002/aic.16367
Abernathy, Mary H., Zhang, Yuchen, Hollinshead, Whitney D., Wang, George, Baidoo, Edward E. K., Liu, Tiangang, and Tang, Yinjie J. Fri . "Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes". United States. https://doi.org/10.1002/aic.16367.
@article{osti_1472164,
title = {Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes},
author = {Abernathy, Mary H. and Zhang, Yuchen and Hollinshead, Whitney D. and Wang, George and Baidoo, Edward E. K. and Liu, Tiangang and Tang, Yinjie J.},
abstractNote = {This study constructed cell‐free glycolytic enzyme systems and compared them to their in vivo functions in Escherichia coli . Under in vitro conditions, flux regulation followed enzyme concentrations and kinetics. In E. coli, only one of the isozymes of phosphofructokinase (PfkA) and fructose‐bisphosphate aldolase (FbaA) facilitate Embden‐Meyerhof‐Parnas (EMP) flux, but under in vitro assays, these isozymes were interchangeable. Additionally, in vitro introduction of the Entner–Doudoroff (ED) pathway improved glycolysis rates, while in vivo overexpression of the ED pathway could not capture significant flux unless its phosphotransferase system (PTS) was knocked out. Lastly, in vivo dynamic 13 C‐experiments revealed that the labeling order of EMP pathway intermediates was not strictly cascade, indicating intracellular metabolites were not well mixed. These enigmatic observations cannot be fully explained by thermodynamics or substrate level regulations. This article supports the long‐time conjecture that EMP enzymes are channeled, and the PTS may be an anchor point to initiate enzyme assemblies. © 2018 American Institute of Chemical Engineers AIChE J , 65: 483–490, 2019},
doi = {10.1002/aic.16367},
journal = {AIChE Journal},
number = 2,
volume = 65,
place = {United States},
year = {Fri Sep 21 00:00:00 EDT 2018},
month = {Fri Sep 21 00:00:00 EDT 2018}
}

Journal Article:
Free Publicly Available Full Text
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https://doi.org/10.1002/aic.16367

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Cited by: 8 works
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