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

doi:10.1016/j.ymben.2017.11.016

Title: 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

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. As a result, 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.

Authors:

Bennett, R. Kyle ^[1]; Gonzalez, Jacqueline E. ^[1]; Whitaker, W. Brian ^[1]; Antoniewicz, Maciek R. ^[1]; Papoutsakis, Eleftherios T. ^[1]

Univ. of Delaware, Newark, DE (United States)

Publication Date:: 2017-12-05

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

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

OSTI Identifier:: 1411402

Grant/Contract Number:: AR0000432

Resource Type:: Accepted Manuscript

Journal Name:: Metabolic Engineering

Additional Journal Information:: Journal Volume: 45; Journal Issue: C; Journal ID: ISSN 1096-7176

Publisher:: Elsevier

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Synthetic methylotrophy; Methanol; Escherichia coli; Pentose phosphate pathway; Phosphoglucose isomerase

Citation Formats


                    Bennett, R. Kyle, Gonzalez, Jacqueline E., Whitaker, W. Brian, Antoniewicz, Maciek R., and Papoutsakis, Eleftherios T. 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.  United States: N. p., 2017. 
        Web.  doi:10.1016/j.ymben.2017.11.016.

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                    Bennett, R. Kyle, Gonzalez, Jacqueline E., Whitaker, W. Brian, Antoniewicz, Maciek R., & Papoutsakis, Eleftherios T. 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.  United States.  doi:10.1016/j.ymben.2017.11.016.

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                    Bennett, R. Kyle, Gonzalez, Jacqueline E., Whitaker, W. Brian, Antoniewicz, Maciek R., and Papoutsakis, Eleftherios T. Tue .  
        "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".  United States.  doi:10.1016/j.ymben.2017.11.016.  https://www.osti.gov/servlets/purl/1411402.

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@article{osti_1411402,

  title        = {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},

  author       = {Bennett, R. Kyle and Gonzalez, Jacqueline E. and Whitaker, W. Brian and Antoniewicz, Maciek R. and Papoutsakis, Eleftherios T.},

  abstractNote = {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. As a result, 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.},

  doi          = {10.1016/j.ymben.2017.11.016},

  journal      = {Metabolic Engineering},

  number       = C,

  volume       = 45,

  place        = {United States},

  year         = {2017},

  month        = {12}

}

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DOI: 10.1016/j.ymben.2017.11.016

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Methanol fermentation increases the production of NAD(P)H-dependent chemicals in synthetic methylotrophic Escherichia coli
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Wang, Xin; Wang, Xuelin; Lu, Xiaolu
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Title: 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

Abstract

Citation Formats

Methanol fermentation increases the production of NAD(P)H-dependent chemicals in synthetic methylotrophic Escherichia coli journal, January 2019

Methanol fermentation increases the production of NAD(P)H-dependent chemicals in synthetic methylotrophic Escherichia coli
journal, January 2019