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

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

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

Journal Article · 05 December 2017 · Metabolic Engineering

DOI: https://doi.org/10.1016/j.ymben.2017.11.016 · OSTI ID:1411402

Bennett, R. Kyle ^[1]; Gonzalez, Jacqueline E. ^[2]; Whitaker, W. Brian ^[2]; Antoniewicz, Maciek R. ^[2]; Papoutsakis, Eleftherios T. ^[2]

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

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.

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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:: 1411402

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Journal Issue: C Vol. 45; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
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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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