Expression of heterologous non-oxidative pentose phosphate pathway from Bacillus methanolicus and phosphoglucose isomerase deletion improves methanol assimilation and metabolite production by a synthetic Escherichia coli methylotroph

R Kyle Bennett¹, Jacqueline E Gonzalez², W Brian Whitaker³, Maciek R Antoniewicz⁴, Eleftherios T Papoutsakis⁵

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: rkbenne@udel.edu.
² Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA. Electronic address: jgonz@udel.edu.
³ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: wbwhit@udel.edu.
⁴ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA. Electronic address: mranton@udel.edu.
⁵ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: epaps@udel.edu.

PMID: 29203223
DOI: 10.1016/j.ymben.2017.11.016

Expression of heterologous non-oxidative pentose phosphate pathway from Bacillus methanolicus and phosphoglucose isomerase deletion improves methanol assimilation and metabolite production by a synthetic Escherichia coli methylotroph

R Kyle Bennett et al. Metab Eng. 2018 Jan.

. 2018 Jan:45:75-85.

doi: 10.1016/j.ymben.2017.11.016. Epub 2017 Dec 5.

Authors

R Kyle Bennett¹, Jacqueline E Gonzalez², W Brian Whitaker³, Maciek R Antoniewicz⁴, Eleftherios T Papoutsakis⁵

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: rkbenne@udel.edu.
² Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA. Electronic address: jgonz@udel.edu.
³ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: wbwhit@udel.edu.
⁴ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA. Electronic address: mranton@udel.edu.
⁵ Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy St., Newark, DE 19716, USA; The Delaware Biotechnology Institute, Molecular Biotechnology Laboratory, University of Delaware, 15 Innovation Way, Newark, DE 19711, USA. Electronic address: epaps@udel.edu.

PMID: 29203223
DOI: 10.1016/j.ymben.2017.11.016

Abstract

Synthetic methylotrophy aims to develop non-native methylotrophic microorganisms to utilize methane or methanol to produce chemicals and biofuels. We report two complimentary strategies to further engineer a previously engineered methylotrophic E. coli strain for improved methanol utilization. First, we demonstrate improved methanol assimilation in the presence of small amounts of yeast extract by expressing the non-oxidative pentose phosphate pathway (PPP) from Bacillus methanolicus. Second, we demonstrate improved co-utilization of methanol and glucose by deleting the phosphoglucose isomerase gene (pgi), which rerouted glucose carbon flux through the oxidative PPP. Both strategies led to significant improvements in methanol assimilation as determined by ¹³C-labeling in intracellular metabolites. Introduction of an acetone-formation pathway in the pgi-deficient methylotrophic E. coli strain led to improved methanol utilization and acetone titers during glucose fed-batch fermentation.

Keywords: Escherichia coli; Methanol; Pentose phosphate pathway; Phosphoglucose isomerase; Synthetic methylotrophy.

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Expression of heterologous non-oxidative pentose phosphate pathway from Bacillus methanolicus and phosphoglucose isomerase deletion improves methanol assimilation and metabolite production by a synthetic Escherichia coli methylotroph

Affiliations

Expression of heterologous non-oxidative pentose phosphate pathway from Bacillus methanolicus and phosphoglucose isomerase deletion improves methanol assimilation and metabolite production by a synthetic Escherichia coli methylotroph

Authors

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Abstract

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