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];
[2];
- Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.
- 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 = {Tue Apr 28 00:00:00 EDT 2020},
month = {Tue Apr 28 00:00:00 EDT 2020}
}
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