Influence of C4-dicarboxylic acid transporters on succinate production

Beauprez, Joeri; Foulquié-Moreno, María R.; Maertens, Jo; Horen, Ellen Van; Bekers, Katelijne M.; Baart, Gino; Cunin, Raymond; Charlier, Daniël; Heijnen, Joseph J.; Soetaert, Wim

doi:10.1039/c1gc15369b

Cited by 11 publications

(9 citation statements)

References 24 publications

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“…Fumarate uptake by E. coli was partly mediated by a dicarboxylic acid transporter that resulted in succinate secretion. Utilization of a dicarboxylic acid transporter for fumarate uptake that is accompanied by succinate secretion has been previously documented in E. coli in certain media [71]. Meanwhile, acetate cross-feeding from E. coli to B. subtilis allowed a larger carbon flow into the TCA cycle in B. subtilis , resulting not only in more energy but also in more 2-oxaloglutarate production by B. subtilis .…”

Section: Resultsmentioning

confidence: 95%

Emergent Biosynthetic Capacity in Simple Microbial Communities

2014

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Microbes have an astonishing capacity to transform their environments. Yet, the metabolic capacity of a single species is limited and the vast majority of microorganisms form complex communities and join forces to exhibit capabilities far exceeding those achieved by any single species. Such enhanced metabolic capacities represent a promising route to many medical, environmental, and industrial applications and call for the development of a predictive, systems-level understanding of synergistic microbial capacity. Here we present a comprehensive computational framework, integrating high-quality metabolic models of multiple species, temporal dynamics, and flux variability analysis, to study the metabolic capacity and dynamics of simple two-species microbial ecosystems. We specifically focus on detecting emergent biosynthetic capacity – instances in which a community growing on some medium produces and secretes metabolites that are not secreted by any member species when growing in isolation on that same medium. Using this framework to model a large collection of two-species communities on multiple media, we demonstrate that emergent biosynthetic capacity is highly prevalent. We identify commonly observed emergent metabolites and metabolic reprogramming patterns, characterizing typical mechanisms of emergent capacity. We further find that emergent secretion tends to occur in two waves, the first as soon as the two organisms are introduced, and the second when the medium is depleted and nutrients become limited. Finally, aiming to identify global community determinants of emergent capacity, we find a marked association between the level of emergent biosynthetic capacity and the functional/phylogenetic distance between community members. Specifically, we demonstrate a “Goldilocks” principle, where high levels of emergent capacity are observed when the species comprising the community are functionally neither too close, nor too distant. Taken together, our results demonstrate the potential to design and engineer synthetic communities capable of novel metabolic activities and point to promising future directions in environmental and clinical bioengineering.

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Section: Resultsmentioning

confidence: 95%

Emergent Biosynthetic Capacity in Simple Microbial Communities

2014

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“…3D), which was consistent with previous results. 30 Moreover, the FA yields of strains A-dcuB-Ec and A-dcuC-Ec were increased by 48.5% and 53.1%, respectively, in 36 hours relative to that of the control strain ABCDIA (Fig. 3C).…”

Section: The Effect Of the Overexpressed Transportersmentioning

confidence: 86%

Regulating C4-dicarboxylate transporters for improving fumaric acid production

et al. 2017

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a Although many efforts have been made to engineer Escherichia coli for fumaric acid production, the fumarate efflux system has not been investigated as an engineering target to improve fumaric acid production. In this work, we cloned and expressed C4-dicarboxylate transporters of different sources in a previously constructed fumaric-acid-producing strain to study their effects on the production of fumaric acid. In addition, each native C4-dicarboxylate transporter was deleted in separate experiments to investigate their individual effects on fumaric acid production. The results showed that the expression of the genes dcuB-Ec and dcuC-Ec can increase the fumaric acid yield by 48.5% and 53.1%, respectively.Fed-batch cultivations in a 5 L bioreactor of strain A-dcuB-Ec produced 9.42 g L À1 of fumaric acid after 50 hours.

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“…This indicates the importance of a more efficient product exporter. This prominence of examining transport processes, such as substrate uptake and product export, possibly coupled to cellular energetics, for microbial organic acid production was mentioned for various organic acids in different host organisms (Beauprez et al, ; Sandström et al, ; Sauer et al, ). Therefore, an even more powerful succinate exporter and the elimination of residual succinate import activity should be the first new targets, because it has been shown that central metabolism is flexible enough to accommodate large changes in fluxes.…”

Section: Resultsmentioning

confidence: 99%

Comparative fluxome and metabolome analysis for overproduction of succinate in Escherichia coli

Taymaz-Nikerel

Mey

Baart

et al. 2015

Biotech & Bioengineering

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An aerobic succinate-producing Escherichia coli mutant was compared to its wild-type by quantitatively analyzing both the metabolome and fluxome, during glucose-limited steady-state and succinate excess dynamic conditions, in order to identify targets for further strain engineering towards more efficient succinate production. The mutant had four functional mutations under the conditions investigated: increased expression of a succinate exporter (DcuC), deletion of a succinate importer (Dct), deletion of succinate dehydrogenase (SUCDH) and expression of a PEP carboxylase (PPC) with increased capacity due to a point mutation. The steady-state and dynamic patterns of the intracellular metabolite levels and fluxes in response to changes were used to locate the quantitative differences in the physiology/metabolism of the mutant strain. Unexpectedly the mutant had a higher energy efficiency, indicated by a much lower rate of oxygen consumption, under glucose-limited conditions, caused by the deletion of the transcription factors IclR and ArcA. Furthermore the mutant had a much lower uptake capacity for succinate (26-fold) and oxygen (17-fold under succinate excess) compared to the wild-type strain. The mutant strain produced 7.9 mmol.CmolX(-1).h(-1) succinate during chemostat cultivation, showing that the choice of the applied genetic modifications was a successful strategy. Furthermore, the applied genetic modifications resulted in multiple large changes in metabolite levels (FBP, pyruvate, 6PG, NAD(+) /NADH ratio, α-ketogluarate) corresponding to large changes in fluxes. Compared to the wild-type a considerable flux shift occurred from the tricarboxylic acid (TCA) cycle to the oxidative part of the pentose phosphate pathway, including an inversion of the pyruvate kinase flux. The mutant responded very differently to excess of succinate, with a remarkable possible reversal of the TCA cycle. The mutant and the wild-type both showed homeostatic behaviour with respect to the energy charge. In contrast, large changes in redox ratios (NAD(+) /NADH) occurred in the wild-type, while the mutant showed even larger changes. This large redox change can be associated to the reversal of flux directions. The observed large flexibility in the central metabolism following genetic (deletions) and environmental (substrate excess) perturbations of the mutant, indicates that introducing a more efficient succinate exporter could result in an even higher succinate production rate.

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Influence of C4-dicarboxylic acid transporters on succinate production

Cited by 11 publications

References 24 publications

Emergent Biosynthetic Capacity in Simple Microbial Communities

Emergent Biosynthetic Capacity in Simple Microbial Communities

Regulating C4-dicarboxylate transporters for improving fumaric acid production

Comparative fluxome and metabolome analysis for overproduction of succinate in Escherichia coli

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