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Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway

BACKGROUND: ʟ-Ornithine is an important amino acid with broad applications in pharmaceutical and food industries. Despite lagging ʟ-ornithine productivity and cost reduction, microbial fermentation is a promising route for sustainable ʟ-ornithine production and thus development of robust microbial s...

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Autores principales: Zhang, Bin, Yu, Miao, Wei, Wen-Ping, Ye, Bang-Ce
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6001011/
https://www.ncbi.nlm.nih.gov/pubmed/29898721
http://dx.doi.org/10.1186/s12934-018-0940-9
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author Zhang, Bin
Yu, Miao
Wei, Wen-Ping
Ye, Bang-Ce
author_facet Zhang, Bin
Yu, Miao
Wei, Wen-Ping
Ye, Bang-Ce
author_sort Zhang, Bin
collection PubMed
description BACKGROUND: ʟ-Ornithine is an important amino acid with broad applications in pharmaceutical and food industries. Despite lagging ʟ-ornithine productivity and cost reduction, microbial fermentation is a promising route for sustainable ʟ-ornithine production and thus development of robust microbial strains with high stability and productivity is essential. RESULTS: Previously, we systematically developed a new strain, SO1 originate from Corynebacterium glutamicum S9114, for ʟ-ornithine production. In this work, overexpression of cg3035 encoding N-acetylglutamate synthase (NAGS) using a plasmid or by inserting a strong P(tac) promoter into the chromosome was found to increase ʟ-ornithine production in the engineered C. glutamicum SO1. The genome-based cg3035 modulated strain was further engineered by attenuating the expression of pta and cat, inserting a strong P(eftu) promoter in the upstream region of glycolytic enzymes such as pfkA, gap, and pyk, and redirecting carbon flux to the pentose phosphate pathway. The final strain with all the exploratory metabolic engineering manipulations produced 32.3 g/L of ʟ-ornithine, a yield of 0.395 g ornithine per g glucose, which was 35.7% higher than that produced by the original strain (23.8 g/L). CONCLUSION: These results clearly demonstrated that enhancing the expression of NAGS promoted ʟ-ornithine production and provide a promising alternative systematic blueprint for developing ʟ-ornithine-producing C. glutamicum strains. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0940-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-60010112018-06-26 Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway Zhang, Bin Yu, Miao Wei, Wen-Ping Ye, Bang-Ce Microb Cell Fact Research BACKGROUND: ʟ-Ornithine is an important amino acid with broad applications in pharmaceutical and food industries. Despite lagging ʟ-ornithine productivity and cost reduction, microbial fermentation is a promising route for sustainable ʟ-ornithine production and thus development of robust microbial strains with high stability and productivity is essential. RESULTS: Previously, we systematically developed a new strain, SO1 originate from Corynebacterium glutamicum S9114, for ʟ-ornithine production. In this work, overexpression of cg3035 encoding N-acetylglutamate synthase (NAGS) using a plasmid or by inserting a strong P(tac) promoter into the chromosome was found to increase ʟ-ornithine production in the engineered C. glutamicum SO1. The genome-based cg3035 modulated strain was further engineered by attenuating the expression of pta and cat, inserting a strong P(eftu) promoter in the upstream region of glycolytic enzymes such as pfkA, gap, and pyk, and redirecting carbon flux to the pentose phosphate pathway. The final strain with all the exploratory metabolic engineering manipulations produced 32.3 g/L of ʟ-ornithine, a yield of 0.395 g ornithine per g glucose, which was 35.7% higher than that produced by the original strain (23.8 g/L). CONCLUSION: These results clearly demonstrated that enhancing the expression of NAGS promoted ʟ-ornithine production and provide a promising alternative systematic blueprint for developing ʟ-ornithine-producing C. glutamicum strains. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0940-9) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-13 /pmc/articles/PMC6001011/ /pubmed/29898721 http://dx.doi.org/10.1186/s12934-018-0940-9 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
Zhang, Bin
Yu, Miao
Wei, Wen-Ping
Ye, Bang-Ce
Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title_full Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title_fullStr Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title_full_unstemmed Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title_short Optimization of ʟ-ornithine production in recombinant Corynebacterium glutamicum S9114 by cg3035 overexpression and manipulating the central metabolic pathway
title_sort optimization of ʟ-ornithine production in recombinant corynebacterium glutamicum s9114 by cg3035 overexpression and manipulating the central metabolic pathway
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6001011/
https://www.ncbi.nlm.nih.gov/pubmed/29898721
http://dx.doi.org/10.1186/s12934-018-0940-9
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