skip to main content

DOE PAGESDOE PAGES

Title: Elucidating the potential of crude cell extracts for producing pyruvate from glucose

Here, living systems possess a rich biochemistry that can be harnessed through metabolic engineering to produce valuable therapeutics, fuels and fine chemicals. In spite of the tools created for this purpose, many organisms tend to be recalcitrant to modification or difficult to optimize. Crude cellular extracts, made by lysis of cells, possess much of the same biochemical capability, but in an easier to manipulate context. Metabolic engineering in crude extracts, or cell-free metabolic engineering, can harness these capabilities to feed heterologous pathways for metabolite production and serve as a platform for pathway optimization. However, the inherent biochemical potential of a crude extract remains ill-defined, and consequently, the use of such extracts can result in inefficient processes and unintended side products. Herein, we show that changes in cell growth conditions lead to changes in the enzymatic activity of crude cell extracts and result in different abilities to produce the central biochemical precursor pyruvate when fed glucose. Proteomic analyses coupled with metabolite measurements uncover the diverse biochemical capabilities of these different crude extract preparations and provide a framework for how analytical measurements can be used to inform and improve crude extract performance. Such informed developments can allow enrichment of crude extracts withmore » pathways that promote or deplete particular metabolic processes and aid in the metabolic engineering of defined products.« less
Authors:
 [1] ;  [1] ;  [2] ; ORCiD logo [2] ; ORCiD logo [2] ; ORCiD logo [1]
  1. Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Publication Date:
Grant/Contract Number:
AC05-00OR22725
Type:
Published Article
Journal Name:
Synthetic Biology
Additional Journal Information:
Journal Volume: 3; Journal Issue: 1; Journal ID: ISSN 2397-7000
Publisher:
Oxford University Press
Research Org:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (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:
1441091
Alternate Identifier(s):
OSTI ID: 1468228

Garcia, David C., Mohr, Benjamin P., Dovgan, Jakob T., Hurst, Gregory B., Standaert, Robert F., and Doktycz, Mitchel John. Elucidating the potential of crude cell extracts for producing pyruvate from glucose. United States: N. p., Web. doi:10.1093/synbio/ysy006.
Garcia, David C., Mohr, Benjamin P., Dovgan, Jakob T., Hurst, Gregory B., Standaert, Robert F., & Doktycz, Mitchel John. Elucidating the potential of crude cell extracts for producing pyruvate from glucose. United States. doi:10.1093/synbio/ysy006.
Garcia, David C., Mohr, Benjamin P., Dovgan, Jakob T., Hurst, Gregory B., Standaert, Robert F., and Doktycz, Mitchel John. 2018. "Elucidating the potential of crude cell extracts for producing pyruvate from glucose". United States. doi:10.1093/synbio/ysy006.
@article{osti_1441091,
title = {Elucidating the potential of crude cell extracts for producing pyruvate from glucose},
author = {Garcia, David C. and Mohr, Benjamin P. and Dovgan, Jakob T. and Hurst, Gregory B. and Standaert, Robert F. and Doktycz, Mitchel John},
abstractNote = {Here, living systems possess a rich biochemistry that can be harnessed through metabolic engineering to produce valuable therapeutics, fuels and fine chemicals. In spite of the tools created for this purpose, many organisms tend to be recalcitrant to modification or difficult to optimize. Crude cellular extracts, made by lysis of cells, possess much of the same biochemical capability, but in an easier to manipulate context. Metabolic engineering in crude extracts, or cell-free metabolic engineering, can harness these capabilities to feed heterologous pathways for metabolite production and serve as a platform for pathway optimization. However, the inherent biochemical potential of a crude extract remains ill-defined, and consequently, the use of such extracts can result in inefficient processes and unintended side products. Herein, we show that changes in cell growth conditions lead to changes in the enzymatic activity of crude cell extracts and result in different abilities to produce the central biochemical precursor pyruvate when fed glucose. Proteomic analyses coupled with metabolite measurements uncover the diverse biochemical capabilities of these different crude extract preparations and provide a framework for how analytical measurements can be used to inform and improve crude extract performance. Such informed developments can allow enrichment of crude extracts with pathways that promote or deplete particular metabolic processes and aid in the metabolic engineering of defined products.},
doi = {10.1093/synbio/ysy006},
journal = {Synthetic Biology},
number = 1,
volume = 3,
place = {United States},
year = {2018},
month = {5}
}