Engineering the biological conversion of methanol to specialty chemicals in Escherichia coli

Whitaker, W. Brian; Jones, J. Andrew; Bennett, R. Kyle; Gonzalez, Jacqueline E.; Vernacchio, Victoria R.; Collins, Shannon M.; Palmer, Michael A.; Schmidt, Samuel; Antoniewicz, Maciek R.; Koffas, Mattheos A.; Papoutsakis, Eleftherios T.

doi:10.1016/j.ymben.2016.10.015

Engineering the biological conversion of methanol to specialty chemicals in Escherichia coli

Journal Article · Tue Nov 01 00:00:00 EDT 2016 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2016.10.015· OSTI ID:1410784

Whitaker, W. Brian ^[1]; Jones, J. Andrew ^[2]; Bennett, R. Kyle ^[3]; Gonzalez, Jacqueline E. ^[4]; Vernacchio, Victoria R. ^[5]; Collins, Shannon M. ^[5]; Palmer, Michael A. ^[4]; Schmidt, Samuel ^[4]; Antoniewicz, Maciek R. ^[4]; Koffas, Mattheos A. ^[6]; Papoutsakis, Eleftherios T. ^[3]

Univ. of Delaware, Newark, DE (United States). Dept. of Chemical and Biomolecular Engineering; Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.; University of Delaware
Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies; Hamilton College, Clinton, NY (United States)
Univ. of Delaware, Newark, DE (United States). Dept. of Chemical and Biomolecular Engineering; Univ. of Delaware, Newark, DE (United States). Delaware Biotechnology Inst.
Univ. of Delaware, Newark, DE (United States). Dept. of Chemical and Biomolecular Engineering
Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies
Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies; Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Biological Sciences, Center for Biotechnology and Interdisciplinary Studies

Methanol is an attractive substrate for biological production of chemicals and fuels. Engineering methylotrophic Escherichia coli as a platform organism for converting methanol to metabolites is desirable. Prior efforts to engineer methylotrophic E. coli were limited by methanol dehydrogenases (Mdhs) with unfavorable enzyme kinetics. We engineered E. coli to utilize methanol using a superior NAD-dependent Mdh from Bacillus stearothermophilus and ribulose monophosphate (RuMP) pathway enzymes from B. methanolicus. Using ¹³C-labeling, we demonstrate this E. coli strain converts methanol into biomass components. For example, the key TCA cycle intermediates, succinate and malate, exhibit labeling up to 39%, while the lower glycolytic intermediate, 3-phosphoglycerate, up to 53%. Multiple carbons are labeled for each compound, demonstrating a cycling RuMP pathway for methanol assimilation to support growth. In conclusion, by incorporating the pathway to synthesize the flavanone naringenin, we demonstrate the first example of in vivo conversion of methanol into a specialty chemical in E. coli.

View Accepted Manuscript (DOE)

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:: 1410784

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Journal Issue: C Vol. 39; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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