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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

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:
 [1] ;  [1] ;  [1] ;  [1] ;  [1]
  1. Univ. of Delaware, Newark, DE (United States)
Publication Date:
Grant/Contract Number:
AR0000432
Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 45; Journal Issue: C; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Research Org:
Univ. of Delaware, Newark, DE (United States)
Sponsoring Org:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
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
OSTI Identifier:
1411402

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., Web. doi:10.1016/j.ymben.2017.11.016.
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.
Bennett, R. Kyle, Gonzalez, Jacqueline E., Whitaker, W. Brian, Antoniewicz, Maciek R., and Papoutsakis, Eleftherios T.. 2017. "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.
@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}
}