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Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

BACKGROUND: Succinate has been recognized as one of the most important bio-based building block chemicals due to its numerous potential applications. However, efficient methods for the production of succinate from lignocellulosic feedstock were rarely reported. Nevertheless, Corynebacterium glutamic...

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Autores principales: Mao, Yufeng, Li, Guiying, Chang, Zhishuai, Tao, Ran, Cui, Zhenzhen, Wang, Zhiwen, Tang, Ya-jie, Chen, Tao, Zhao, Xueming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5883316/
https://www.ncbi.nlm.nih.gov/pubmed/29636817
http://dx.doi.org/10.1186/s13068-018-1094-z
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author Mao, Yufeng
Li, Guiying
Chang, Zhishuai
Tao, Ran
Cui, Zhenzhen
Wang, Zhiwen
Tang, Ya-jie
Chen, Tao
Zhao, Xueming
author_facet Mao, Yufeng
Li, Guiying
Chang, Zhishuai
Tao, Ran
Cui, Zhenzhen
Wang, Zhiwen
Tang, Ya-jie
Chen, Tao
Zhao, Xueming
author_sort Mao, Yufeng
collection PubMed
description BACKGROUND: Succinate has been recognized as one of the most important bio-based building block chemicals due to its numerous potential applications. However, efficient methods for the production of succinate from lignocellulosic feedstock were rarely reported. Nevertheless, Corynebacterium glutamicum was engineered to efficiently produce succinate from glucose in our previous study. RESULTS: In this work, C. glutamicum was engineered for efficient succinate production from lignocellulosic hydrolysate. First, xylose utilization of C. glutamicum was optimized by heterologous expression of xylA and xylB genes from different sources. Next, xylA and xylB from Xanthomonas campestris were selected among four candidates to accelerate xylose consumption and cell growth. Subsequently, the optimal xylA and xylB were co-expressed in C. glutamicum strain SAZ3 (ΔldhAΔptaΔpqoΔcatP(sod)-ppcP(sod)-pyc) along with genes encoding pyruvate carboxylase, citrate synthase, and a succinate exporter to achieve succinate production from xylose in a two-stage fermentation process. Xylose utilization and succinate production were further improved by overexpressing the endogenous tkt and tal genes and introducing araE from Bacillus subtilis. The final strain C. glutamicum CGS5 showed an excellent ability to produce succinate in two-stage fermentations by co-utilizing a glucose–xylose mixture under anaerobic conditions. A succinate titer of 98.6 g L(−1) was produced from corn stalk hydrolysate with a yield of 0.87 g/g total substrates and a productivity of 4.29 g L(−1) h(−1) during the anaerobic stage. CONCLUSION: This work introduces an efficient process for the bioconversion of biomass into succinate using a thoroughly engineered strain of C. glutamicum. To the best of our knowledge, this is the highest titer of succinate produced from non-food lignocellulosic feedstock, which highlights that the biosafety level 1 microorganism C. glutamicum is a promising platform for the envisioned lignocellulosic biorefinery. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1094-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-58833162018-04-10 Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate Mao, Yufeng Li, Guiying Chang, Zhishuai Tao, Ran Cui, Zhenzhen Wang, Zhiwen Tang, Ya-jie Chen, Tao Zhao, Xueming Biotechnol Biofuels Research BACKGROUND: Succinate has been recognized as one of the most important bio-based building block chemicals due to its numerous potential applications. However, efficient methods for the production of succinate from lignocellulosic feedstock were rarely reported. Nevertheless, Corynebacterium glutamicum was engineered to efficiently produce succinate from glucose in our previous study. RESULTS: In this work, C. glutamicum was engineered for efficient succinate production from lignocellulosic hydrolysate. First, xylose utilization of C. glutamicum was optimized by heterologous expression of xylA and xylB genes from different sources. Next, xylA and xylB from Xanthomonas campestris were selected among four candidates to accelerate xylose consumption and cell growth. Subsequently, the optimal xylA and xylB were co-expressed in C. glutamicum strain SAZ3 (ΔldhAΔptaΔpqoΔcatP(sod)-ppcP(sod)-pyc) along with genes encoding pyruvate carboxylase, citrate synthase, and a succinate exporter to achieve succinate production from xylose in a two-stage fermentation process. Xylose utilization and succinate production were further improved by overexpressing the endogenous tkt and tal genes and introducing araE from Bacillus subtilis. The final strain C. glutamicum CGS5 showed an excellent ability to produce succinate in two-stage fermentations by co-utilizing a glucose–xylose mixture under anaerobic conditions. A succinate titer of 98.6 g L(−1) was produced from corn stalk hydrolysate with a yield of 0.87 g/g total substrates and a productivity of 4.29 g L(−1) h(−1) during the anaerobic stage. CONCLUSION: This work introduces an efficient process for the bioconversion of biomass into succinate using a thoroughly engineered strain of C. glutamicum. To the best of our knowledge, this is the highest titer of succinate produced from non-food lignocellulosic feedstock, which highlights that the biosafety level 1 microorganism C. glutamicum is a promising platform for the envisioned lignocellulosic biorefinery. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1094-z) contains supplementary material, which is available to authorized users. BioMed Central 2018-04-04 /pmc/articles/PMC5883316/ /pubmed/29636817 http://dx.doi.org/10.1186/s13068-018-1094-z 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
Mao, Yufeng
Li, Guiying
Chang, Zhishuai
Tao, Ran
Cui, Zhenzhen
Wang, Zhiwen
Tang, Ya-jie
Chen, Tao
Zhao, Xueming
Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title_full Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title_fullStr Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title_full_unstemmed Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title_short Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
title_sort metabolic engineering of corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5883316/
https://www.ncbi.nlm.nih.gov/pubmed/29636817
http://dx.doi.org/10.1186/s13068-018-1094-z
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