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A metabolic reconstruction of Lactobacillus reuteri JCM 1112 and analysis of its potential as a cell factory

A metabolic reconstruction of Lactobacillus reuteri JCM 1112 and analysis of its potential as a cell factory


Title: A metabolic reconstruction of Lactobacillus reuteri JCM 1112 and analysis of its potential as a cell factory
Author: Kristjánsdóttir, Þórdís   orcid.org/0000-0003-2244-1723
Bosma, Elleke F.
Branco dos Santos, Filipe
Özdemir, Emre
Herrgård, Markus J.
França, Lucas
Ferreira, Bruno
Nielsen, Alex T.
Guðmundsson, Steinn   orcid.org/0000-0002-2758-2720
Date: 2019-10-29
Language: English
Scope: 186
University/Institute: Háskóli Íslands
University of Iceland
School: School of Engineering and Natural Sciences (UI)
Verkfræði- og náttúruvísindasvið (HÍ)
Department: Center for Systems Biology (UI)
Rannsóknarsetur í kerfislíffræði (HÍ)
Series: Microbial Cell Factories;18(1)
ISSN: 1475-2859
DOI: 10.1186/s12934-019-1229-3
Subject: Cell factory; Genome-scale metabolic model; Lactobacillus reuteri; Gerlar; Frumulíffræði; Genamengi
URI: https://hdl.handle.net/20.500.11815/1611

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Citation:

Kristjansdottir, T., Bosma, E.F., Branco dos Santos, F. et al. A metabolic reconstruction of Lactobacillus reuteri JCM 1112 and analysis of its potential as a cell factory. Microb Cell Fact 18, 186 (2019). https://doi.org/10.1186/s12934-019-1229-3

Abstract:

Background: Lactobacillus reuteri is a heterofermentative Lactic Acid Bacterium (LAB) that is commonly used for food fermentations and probiotic purposes. Due to its robust properties, it is also increasingly considered for use as a cell factory. It produces several industrially important compounds such as 1,3-propanediol and reuterin natively, but for cell factory purposes, developing improved strategies for engineering and fermentation optimization is crucial. Genome-scale metabolic models can be highly beneficial in guiding rational metabolic engineering. Reconstructing a reliable and a quantitatively accurate metabolic model requires extensive manual curation and incorporation of experimental data. Results: A genome-scale metabolic model of L. reuteri JCM 1112T was reconstructed and the resulting model, Lreuteri_530, was validated and tested with experimental data. Several knowledge gaps in the metabolism were identified and resolved during this process, including presence/absence of glycolytic genes. Flux distribution between the two glycolytic pathways, the phosphoketolase and Embden-Meyerhof-Parnas pathways, varies considerably between LAB species and strains. As these pathways result in different energy yields, it is important to include strain-specific utilization of these pathways in the model. We determined experimentally that the Embden-Meyerhof-Parnas pathway carried at most 7% of the total glycolytic flux. Predicted growth rates from Lreuteri_530 were in good agreement with experimentally determined values. To further validate the prediction accuracy of Lreuteri_530, the predicted effects of glycerol addition and adhE gene knock-out, which results in impaired ethanol production, were compared to in vivo data. Examination of both growth rates and uptake- and secretion rates of the main metabolites in central metabolism demonstrated that the model was able to accurately predict the experimentally observed effects. Lastly, the potential of L. reuteri as a cell factory was investigated, resulting in a number of general metabolic engineering strategies. Conclusion: We have constructed a manually curated genome-scale metabolic model of L. reuteri JCM 1112T that has been experimentally parameterized and validated and can accurately predict metabolic behavior of this important platform cell factory.

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Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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