Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli

Bennett, R. Kyle; Agee, Alec; Gerald Har, Jie Ren; von Hagel, Bryan; Siu, Ka-Hei; Antoniewicz, Maciek R.; Papoutsakis, Eleftherios T.

doi:10.1016/j.ymben.2020.04.007

Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli

Journal Article · Tue Apr 28 00:00:00 EDT 2020 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2020.04.007· OSTI ID:1742083

Bennett, R. Kyle ^[1]; ^[2]; ^[3]; von Hagel, Bryan ^[2]; Siu, Ka-Hei ^[2]; Antoniewicz, Maciek R. ^[3]; Papoutsakis, Eleftherios T. ^[2]

Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.; University of Delaware
Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.
Univ. of Delaware, Newark, DE (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Delaware, Newark, DE (United States)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E)

Grant/Contract Number:: AR0000432

OSTI ID:: 1742083

Alternate ID(s):: OSTI ID: 1617240

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Vol. 61; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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