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Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum

BACKGROUND: α-d-Glucosylglycerol (αGG) has beneficial functions as a moisturizing agent in cosmetics and potential as a health food material, and therapeutic agent. αGG serves as compatible solute in various halotolerant cyanobacteria such as Synechocystis sp. PCC 6803, which synthesizes αGG in a tw...

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Autores principales: Roenneke, Benjamin, Rosenfeldt, Natalie, Derya, Sami M., Novak, Jens F., Marin, Kay, Krämer, Reinhard, Seibold, Gerd M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6004087/
https://www.ncbi.nlm.nih.gov/pubmed/29908566
http://dx.doi.org/10.1186/s12934-018-0939-2
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author Roenneke, Benjamin
Rosenfeldt, Natalie
Derya, Sami M.
Novak, Jens F.
Marin, Kay
Krämer, Reinhard
Seibold, Gerd M.
author_facet Roenneke, Benjamin
Rosenfeldt, Natalie
Derya, Sami M.
Novak, Jens F.
Marin, Kay
Krämer, Reinhard
Seibold, Gerd M.
author_sort Roenneke, Benjamin
collection PubMed
description BACKGROUND: α-d-Glucosylglycerol (αGG) has beneficial functions as a moisturizing agent in cosmetics and potential as a health food material, and therapeutic agent. αGG serves as compatible solute in various halotolerant cyanobacteria such as Synechocystis sp. PCC 6803, which synthesizes αGG in a two-step reaction: The enzymatic condensation of ADP-glucose and glycerol 3-phosphate by GG-phosphate synthase (GGPS) is followed by the dephosphorylation of the intermediate by the GG-phosphate phosphatase (GGPP). The Gram-positive Corynebacterium glutamicum, an industrial workhorse for amino acid production, does not utilize αGG as a substrate and was therefore chosen for the development of a heterologous microbial production platform for αGG. RESULTS: Plasmid-bound expression of ggpS and ggpP from Synechocystis sp. PCC 6803 enabled αGG synthesis exclusively in osmotically stressed cells of C. glutamicum (pEKEx2-ggpSP), which is probably due to the unique intrinsic control mechanism of GGPS activity in response to intracellular ion concentrations. C. glutamicum was then engineered to optimize precursor supply for αGG production: The precursor for αGG synthesis ADP-glucose gets metabolized by both the glgA encoded glycogen synthase and the otsA encoded trehalose-6-phosphate synthase. Upon deletion of both genes the αGG concentration in culture supernatants was increased from 0.5 mM in C. glutamicum (pEKEx3-ggpSP) to 2.9 mM in C. glutamicum ΔotsA IMglgA (pEKEx3-ggpSP). Upon nitrogen limitation, which inhibits synthesis of amino acids as compatible solutes, C. glutamicum ΔotsA IMglgA (pEKEx3-ggpSP) produced more than 10 mM αGG (about 2 g L(−1)). CONCLUSIONS: Corynebacterium glutamicum can be engineered as efficient platform for the production of the compatible solute αGG. Redirection of carbon flux towards αGG synthesis by elimination of the competing pathways for glycogen and trehalose synthesis as well as optimization of nitrogen supply is an efficient strategy to further optimize production of αGG. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0939-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-60040872018-06-26 Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum Roenneke, Benjamin Rosenfeldt, Natalie Derya, Sami M. Novak, Jens F. Marin, Kay Krämer, Reinhard Seibold, Gerd M. Microb Cell Fact Research BACKGROUND: α-d-Glucosylglycerol (αGG) has beneficial functions as a moisturizing agent in cosmetics and potential as a health food material, and therapeutic agent. αGG serves as compatible solute in various halotolerant cyanobacteria such as Synechocystis sp. PCC 6803, which synthesizes αGG in a two-step reaction: The enzymatic condensation of ADP-glucose and glycerol 3-phosphate by GG-phosphate synthase (GGPS) is followed by the dephosphorylation of the intermediate by the GG-phosphate phosphatase (GGPP). The Gram-positive Corynebacterium glutamicum, an industrial workhorse for amino acid production, does not utilize αGG as a substrate and was therefore chosen for the development of a heterologous microbial production platform for αGG. RESULTS: Plasmid-bound expression of ggpS and ggpP from Synechocystis sp. PCC 6803 enabled αGG synthesis exclusively in osmotically stressed cells of C. glutamicum (pEKEx2-ggpSP), which is probably due to the unique intrinsic control mechanism of GGPS activity in response to intracellular ion concentrations. C. glutamicum was then engineered to optimize precursor supply for αGG production: The precursor for αGG synthesis ADP-glucose gets metabolized by both the glgA encoded glycogen synthase and the otsA encoded trehalose-6-phosphate synthase. Upon deletion of both genes the αGG concentration in culture supernatants was increased from 0.5 mM in C. glutamicum (pEKEx3-ggpSP) to 2.9 mM in C. glutamicum ΔotsA IMglgA (pEKEx3-ggpSP). Upon nitrogen limitation, which inhibits synthesis of amino acids as compatible solutes, C. glutamicum ΔotsA IMglgA (pEKEx3-ggpSP) produced more than 10 mM αGG (about 2 g L(−1)). CONCLUSIONS: Corynebacterium glutamicum can be engineered as efficient platform for the production of the compatible solute αGG. Redirection of carbon flux towards αGG synthesis by elimination of the competing pathways for glycogen and trehalose synthesis as well as optimization of nitrogen supply is an efficient strategy to further optimize production of αGG. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0939-2) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-16 /pmc/articles/PMC6004087/ /pubmed/29908566 http://dx.doi.org/10.1186/s12934-018-0939-2 Text en © The Author(s) 2018 Open AccessThis 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.
spellingShingle Research
Roenneke, Benjamin
Rosenfeldt, Natalie
Derya, Sami M.
Novak, Jens F.
Marin, Kay
Krämer, Reinhard
Seibold, Gerd M.
Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title_full Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title_fullStr Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title_full_unstemmed Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title_short Production of the compatible solute α-d-glucosylglycerol by metabolically engineered Corynebacterium glutamicum
title_sort production of the compatible solute α-d-glucosylglycerol by metabolically engineered corynebacterium glutamicum
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6004087/
https://www.ncbi.nlm.nih.gov/pubmed/29908566
http://dx.doi.org/10.1186/s12934-018-0939-2
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