Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain

Yinjie J. Tang; Rajat Sapra; Dominique Joyner; Terry C. Hazen; Samuel Myers; David Reichmuth; Harvey Blanch; Jay D. Keasling

doi:10.1002/bit.22181

journal article Nov 17, 2008

Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain

Yinjie J. Tang

Rajat Sapra Dominique Joyner Terry C. Hazen

Samuel Myers David Reichmuth Harvey Blanch Jay D. Keasling

Biotechnology and Bioengineering Vol. 102 No. 5 pp. 1377-1386 · Wiley

View at Publisher Save 10.1002/bit.22181

Abstract

AbstractA recently discovered thermophilic bacterium, Geobacillus thermoglucosidasius M10EXG, ferments a range of C5 (e.g., xylose) and C6 sugars (e.g., glucose) and is tolerant to high ethanol concentrations (10%, v/v). We have investigated the central metabolism of this bacterium using both in vitro enzyme assays and 13C‐based flux analysis to provide insights into the physiological properties of this extremophile and explore its metabolism for bio‐ethanol or other bioprocess applications. Our findings show that glucose metabolism in G. thermoglucosidasius M10EXG proceeds via glycolysis, the pentose phosphate pathway, and the TCA cycle; the Entner–Doudoroff pathway and transhydrogenase activity were not detected. Anaplerotic reactions (including the glyoxylate shunt, pyruvate carboxylase, and phosphoenolpyruvate carboxykinase) were active, but fluxes through those pathways could not be accurately determined using amino acid labeling. When growth conditions were switched from aerobic to micro‐aerobic conditions, fluxes (based on a normalized glucose uptake rate of 100 units (g DCW)−1 h−1) through the TCA cycle and oxidative pentose phosphate pathway were reduced from 64 ± 3 to 25 ± 2 and from 30 ± 2 to 19 ± 2, respectively. The carbon flux under micro‐aerobic growth was directed to ethanol, L‐lactate (>99% optical purity), acetate, and formate. Under fully anerobic conditions, G. thermoglucosidasius M10EXG used a mixed acid fermentation process and exhibited a maximum ethanol yield of 0.38 ± 0.07 mol mol−1 glucose. In silico flux balance modeling demonstrates that lactate and acetate production from G. thermoglucosidasius M10EXG reduces the maximum ethanol yield by approximately threefold, thus indicating that both pathways should be modified to maximize ethanol production. Biotechnol. Bioeng. 2009;102: 1377–1386. © 2008 Wiley Periodicals, Inc.

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Details

Published: Nov 17, 2008
Vol/Issue: 102(5)
Pages: 1377-1386
License: View

Authors

Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA.; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.; Synthetic Biology Engineering Research Center, Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA.

Cite This Article

Yinjie J. Tang, Rajat Sapra, Dominique Joyner, et al. (2008). Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain. Biotechnology and Bioengineering, 102(5), 1377-1386. https://doi.org/10.1002/bit.22181

Analysis of metabolic pathways and fluxes in a newly discovered thermophilic and ethanol‐tolerant Geobacillus strain

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