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Title: A cell-free system for production of 2,3-butanediol is robust to growth-toxic compounds

Abstract

The need for sustainable, low-cost production of bioenergy and commodity chemicals is increasing. Unfortunately, the engineering potential of whole-cell catalysts to address this need can be hampered by cellular toxicity. When such bottlenecks limit the commercial feasibility of whole-cell fermentation, cell-free, or in vitro, based approaches may offer an alternative. Here, we assess the impact of three classes of growth toxic compounds on crude extract-based, cell-free chemical conversions. As a model system, we test a metabolic pathway for conversion of glucose to 2,3-butanediol (2,3-BDO) in lysates of Escherichia coli. First, we characterized 2,3-BDO production with different classes of antibiotics and found, as expected, that the system is uninhibited by compounds that prevent cell growth by means of cell wall replication and DNA, RNA, and protein synthesis. Second, we considered the impact of polar solvent addition (e.g., methanol, n-butanol). We observed that volumetric productivities (g/L/h) were slowed with increasing hydrophobicity of added alcohols. Finally, we investigated the effects of using pretreated biomass hydrolysate as a feed stock, observing a 25% reduction in 2,3-BDO production as a result of coumaroyl and feruloyl amides. Overall, we find the cell-free system to be robust to working concentrations of antibiotics and other compounds that aremore » toxic to cell growth, but do not denature or inhibit relevant enzymes.« less

Authors:
;
Publication Date:
Research Org.:
Northwestern Univ., Evanston, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; David and Lucile Packard Foundation; Dreyfus Teacher-Scholar Program
OSTI Identifier:
1575942
Alternate Identifier(s):
OSTI ID: 1660706
Grant/Contract Number:  
SC0018249
Resource Type:
Published Article
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Name: Metabolic Engineering Communications Journal Volume: 10 Journal Issue: C; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Country of Publication:
Netherlands
Language:
English
Subject:
09 BIOMASS FUELS

Citation Formats

Kay, Jennifer E., and Jewett, Michael C. A cell-free system for production of 2,3-butanediol is robust to growth-toxic compounds. Netherlands: N. p., 2019. Web. doi:10.1016/j.mec.2019.e00114.
Kay, Jennifer E., & Jewett, Michael C. A cell-free system for production of 2,3-butanediol is robust to growth-toxic compounds. Netherlands. https://doi.org/10.1016/j.mec.2019.e00114
Kay, Jennifer E., and Jewett, Michael C. Wed . "A cell-free system for production of 2,3-butanediol is robust to growth-toxic compounds". Netherlands. https://doi.org/10.1016/j.mec.2019.e00114.
@article{osti_1575942,
title = {A cell-free system for production of 2,3-butanediol is robust to growth-toxic compounds},
author = {Kay, Jennifer E. and Jewett, Michael C.},
abstractNote = {The need for sustainable, low-cost production of bioenergy and commodity chemicals is increasing. Unfortunately, the engineering potential of whole-cell catalysts to address this need can be hampered by cellular toxicity. When such bottlenecks limit the commercial feasibility of whole-cell fermentation, cell-free, or in vitro, based approaches may offer an alternative. Here, we assess the impact of three classes of growth toxic compounds on crude extract-based, cell-free chemical conversions. As a model system, we test a metabolic pathway for conversion of glucose to 2,3-butanediol (2,3-BDO) in lysates of Escherichia coli. First, we characterized 2,3-BDO production with different classes of antibiotics and found, as expected, that the system is uninhibited by compounds that prevent cell growth by means of cell wall replication and DNA, RNA, and protein synthesis. Second, we considered the impact of polar solvent addition (e.g., methanol, n-butanol). We observed that volumetric productivities (g/L/h) were slowed with increasing hydrophobicity of added alcohols. Finally, we investigated the effects of using pretreated biomass hydrolysate as a feed stock, observing a 25% reduction in 2,3-BDO production as a result of coumaroyl and feruloyl amides. Overall, we find the cell-free system to be robust to working concentrations of antibiotics and other compounds that are toxic to cell growth, but do not denature or inhibit relevant enzymes.},
doi = {10.1016/j.mec.2019.e00114},
journal = {Metabolic Engineering Communications},
number = C,
volume = 10,
place = {Netherlands},
year = {2019},
month = {11}
}

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