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Title: Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli

Abstract

Synthetic methylotrophy aims to engineer methane and methanol utilization pathways in platform hosts like Escherichia coli for industrial bioprocessing of natural gas and biogas. While recent attempts to engineer synthetic methylotrophs have proved successful, autonomous methylotrophy, i.e. the ability to utilize methane or methanol as sole carbon and energy substrates, has not yet been realized. Here in this paper, we address an important limitation of autonomous methylotrophy in E. coli: the inability of the organism to synthesize several amino acids when grown on methanol. By activating the stringent/stress response via ppGpp overproduction, or DksA and RpoS overexpression, we demonstrate improved biosynthesis of proteinogenic amino acids via endogenous upregulation of amino acid synthesis pathway genes. Thus, we were able to achieve biosynthesis of several limiting amino acids from methanol-derived carbon, in contrast to the control methylotrophic E. coli strain. This study addresses a key limitation currently preventing autonomous methylotrophy in E. coli and possibly other synthetic methylotrophs and provides insight as to how this limitation can be alleviated via stringent/stress response activation.

Authors:
 [1]; ORCiD logo [1]; ORCiD logo [2];  [1];  [1];  [2];  [1]
  1. Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.
  2. Univ. of Delaware, Newark, DE (United States)
Publication Date:
Research Org.:
Univ. of Delaware, Newark, DE (United States)
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1742083
Alternate Identifier(s):
OSTI ID: 1617240
Grant/Contract Number:  
AR0000432
Resource Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 61; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; Synthetic methylotrophy; Methanol; Escherichia coli; Stringent response; ppGpp

Citation Formats

Bennett, R. Kyle, Agee, Alec, Gerald Har, Jie Ren, von Hagel, Bryan, Siu, Ka-Hei, Antoniewicz, Maciek R., and Papoutsakis, Eleftherios T. Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli. United States: N. p., 2020. Web. doi:10.1016/j.ymben.2020.04.007.
Bennett, R. Kyle, Agee, Alec, Gerald Har, Jie Ren, von Hagel, Bryan, Siu, Ka-Hei, Antoniewicz, Maciek R., & Papoutsakis, Eleftherios T. Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli. United States. https://doi.org/10.1016/j.ymben.2020.04.007
Bennett, R. Kyle, Agee, Alec, Gerald Har, Jie Ren, von Hagel, Bryan, Siu, Ka-Hei, Antoniewicz, Maciek R., and Papoutsakis, Eleftherios T. Tue . "Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli". United States. https://doi.org/10.1016/j.ymben.2020.04.007. https://www.osti.gov/servlets/purl/1742083.
@article{osti_1742083,
title = {Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli},
author = {Bennett, R. Kyle and Agee, Alec and Gerald Har, Jie Ren and von Hagel, Bryan and Siu, Ka-Hei and Antoniewicz, Maciek R. and Papoutsakis, Eleftherios T.},
abstractNote = {Synthetic methylotrophy aims to engineer methane and methanol utilization pathways in platform hosts like Escherichia coli for industrial bioprocessing of natural gas and biogas. While recent attempts to engineer synthetic methylotrophs have proved successful, autonomous methylotrophy, i.e. the ability to utilize methane or methanol as sole carbon and energy substrates, has not yet been realized. Here in this paper, we address an important limitation of autonomous methylotrophy in E. coli: the inability of the organism to synthesize several amino acids when grown on methanol. By activating the stringent/stress response via ppGpp overproduction, or DksA and RpoS overexpression, we demonstrate improved biosynthesis of proteinogenic amino acids via endogenous upregulation of amino acid synthesis pathway genes. Thus, we were able to achieve biosynthesis of several limiting amino acids from methanol-derived carbon, in contrast to the control methylotrophic E. coli strain. This study addresses a key limitation currently preventing autonomous methylotrophy in E. coli and possibly other synthetic methylotrophs and provides insight as to how this limitation can be alleviated via stringent/stress response activation.},
doi = {10.1016/j.ymben.2020.04.007},
journal = {Metabolic Engineering},
number = ,
volume = 61,
place = {United States},
year = {2020},
month = {4}
}

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