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

Alternate Identifier(s):: OSTI ID: 1703839

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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Works referencing / citing this record:

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Fan, Liwen; Wang, Yu; Tuyishime, Philibert
ChemBioChem, Vol. 19, Issue 23
DOI: 10.1002/cbic.201800424

Biological conversion of methane to chemicals and fuels: technical challenges and issues
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Applied Microbiology and Biotechnology, Vol. 102, Issue 7
DOI: 10.1007/s00253-018-8842-7

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journal, April 2018

Meyer, Fabian; Keller, Philipp; Hartl, Johannes
Nature Communications, Vol. 9, Issue 1
DOI: 10.1038/s41467-018-03937-y

Improving formaldehyde consumption drives methanol assimilation in engineered E. coli
journal, June 2018

Woolston, Benjamin M.; King, Jason R.; Reiter, Michael
Nature Communications, Vol. 9, Issue 1
DOI: 10.1038/s41467-018-04795-4

Growth of E. coli on formate and methanol via the reductive glycine pathway
journal, February 2020

Kim, Seohyoung; Lindner, Steffen N.; Aslan, Selçuk
Nature Chemical Biology, Vol. 16, Issue 5
DOI: 10.1038/s41589-020-0473-5

C1 Compound Biosensors: Design, Functional Study, and Applications
journal, May 2019

Lee, Jin-Young; Sung, Bong Hyun; Oh, So-Hyung
International Journal of Molecular Sciences, Vol. 20, Issue 9
DOI: 10.3390/ijms20092253

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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

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