Comprehensive analysis of glucose and xylose metabolism in Escherichia coli under aerobic and anaerobic conditions by 13C metabolic flux analysis

Gonzalez, Jacqueline E.; Long, Christopher P.; Antoniewicz, Maciek R.

doi:10.1016/j.ymben.2016.11.003

Title: Comprehensive analysis of glucose and xylose metabolism in Escherichia coli under aerobic and anaerobic conditions by ¹³C metabolic flux analysis

Journal Article · Fri Nov 11 00:00:00 EST 2016 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2016.11.003· OSTI ID:1463265

Gonzalez, Jacqueline E. ^[1]; Long, Christopher P. ^[1]; Antoniewicz, Maciek R. ^[1]

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

Glucose and xylose are the two most abundant sugars derived from the breakdown of lignocellulosic biomass. While aerobic glucose metabolism is relatively well understood in E. coli, until now there have been only a handful of studies focused on anaerobic glucose metabolism and no ¹³C-flux studies on xylose metabolism. In the absence of experimentally validated flux maps, constraint-based approaches such as MOMA and RELATCH cannot be used to guide new metabolic engineering designs. In this work, we have addressed this critical gap in current understanding by performing comprehensive characterizations of glucose and xylose metabolism under aerobic and anaerobic conditions, using recent state-of-the-art techniques in ¹³C metabolic flux analysis (¹³C-MFA). Specifically, we quantified precise metabolic fluxes for each condition by performing parallel labeling experiments and analyzing the data through integrated 13C-MFA using the optimal tracers [1,2-¹³C]glucose, [1,6-¹³C]glucose, [1,2-¹³C]xylose and [5-¹³C]xylose. We also quantified changes in biomass composition and confirmed turnover of macromolecules by applying [U-¹³C]glucose and [U-13C]xylose tracers. We demonstrate that under anaerobic growth conditions there is significant turnover of lipids and that a significant portion of CO₂ originates from biomass turnover. Using knockout strains, we also demonstrate that β-oxidation is critical for anaerobic growth on xylose. Quantitative analysis of co-factor balances (NADH/FADH2, NADPH, and ATP) for different growth conditions provided new insights regarding the interplay of energy and redox metabolism and the impact on E. coli cell physiology.

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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:: 1463265

Alternate ID(s):: OSTI ID: 1397057

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

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 82 works

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