Cargando…
Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution
BACKGROUND: Production directly from carbon dioxide by engineered cyanobacteria is one of the promising technologies for sustainable future. Previously, we have successfully achieved 1,3-propanediol (1,3-PDO) production using Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. The s...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5702090/ https://www.ncbi.nlm.nih.gov/pubmed/29178875 http://dx.doi.org/10.1186/s12934-017-0824-4 |
_version_ | 1783281452169822208 |
---|---|
author | Hirokawa, Yasutaka Matsuo, Shingo Hamada, Hiroyuki Matsuda, Fumio Hanai, Taizo |
author_facet | Hirokawa, Yasutaka Matsuo, Shingo Hamada, Hiroyuki Matsuda, Fumio Hanai, Taizo |
author_sort | Hirokawa, Yasutaka |
collection | PubMed |
description | BACKGROUND: Production directly from carbon dioxide by engineered cyanobacteria is one of the promising technologies for sustainable future. Previously, we have successfully achieved 1,3-propanediol (1,3-PDO) production using Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. The strain into which the synthetic metabolic pathway was introduced produced 3.48 mM (0.265 g/L) 1,3-PDO and 14.3 mM (1.32 g/L) glycerol during 20 days of incubation. In this study, the productivities of 1,3-PDO were improved by gene disruption selected by screening with in silico simulation. METHODS: First, a stoichiometric metabolic model was applied to prediction of cellular metabolic flux distribution in a 1,3-PDO-producing strain of S. elongatus PCC 7942. A genome-scale model of S. elongatus PCC 7942 constructed by Knoop was modified by the addition of a synthetic metabolic pathway for 1,3-PDO production. Next, the metabolic flux distribution predicted by metabolic flux balance analysis (FBA) was used for in silico simulation of gene disruption. As a result of gene disruption simulation, NADPH dehydrogenase 1 (NDH-1) complexes were found by screening to be the most promising candidates for disruption to improve 1,3-PDO production. The effect of disruption of the gene encoding a subunit of the NDH-1 complex was evaluated in the 1,3-PDO-producing strain. RESULTS AND CONCLUSIONS: During 20 days of incubation, the ndhF1-null 1,3-PDO-producing strain showed the highest titers: 4.44 mM (0.338 g/L) 1,3-PDO and 30.3 mM (2.79 g/L) glycerol. In this study, we successfully improved 1,3-PDO productivity on the basis of in silico simulation of gene disruption. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-017-0824-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5702090 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-57020902017-12-04 Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution Hirokawa, Yasutaka Matsuo, Shingo Hamada, Hiroyuki Matsuda, Fumio Hanai, Taizo Microb Cell Fact Research BACKGROUND: Production directly from carbon dioxide by engineered cyanobacteria is one of the promising technologies for sustainable future. Previously, we have successfully achieved 1,3-propanediol (1,3-PDO) production using Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. The strain into which the synthetic metabolic pathway was introduced produced 3.48 mM (0.265 g/L) 1,3-PDO and 14.3 mM (1.32 g/L) glycerol during 20 days of incubation. In this study, the productivities of 1,3-PDO were improved by gene disruption selected by screening with in silico simulation. METHODS: First, a stoichiometric metabolic model was applied to prediction of cellular metabolic flux distribution in a 1,3-PDO-producing strain of S. elongatus PCC 7942. A genome-scale model of S. elongatus PCC 7942 constructed by Knoop was modified by the addition of a synthetic metabolic pathway for 1,3-PDO production. Next, the metabolic flux distribution predicted by metabolic flux balance analysis (FBA) was used for in silico simulation of gene disruption. As a result of gene disruption simulation, NADPH dehydrogenase 1 (NDH-1) complexes were found by screening to be the most promising candidates for disruption to improve 1,3-PDO production. The effect of disruption of the gene encoding a subunit of the NDH-1 complex was evaluated in the 1,3-PDO-producing strain. RESULTS AND CONCLUSIONS: During 20 days of incubation, the ndhF1-null 1,3-PDO-producing strain showed the highest titers: 4.44 mM (0.338 g/L) 1,3-PDO and 30.3 mM (2.79 g/L) glycerol. In this study, we successfully improved 1,3-PDO productivity on the basis of in silico simulation of gene disruption. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-017-0824-4) contains supplementary material, which is available to authorized users. BioMed Central 2017-11-25 /pmc/articles/PMC5702090/ /pubmed/29178875 http://dx.doi.org/10.1186/s12934-017-0824-4 Text en © The Author(s) 2017 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 Hirokawa, Yasutaka Matsuo, Shingo Hamada, Hiroyuki Matsuda, Fumio Hanai, Taizo Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title | Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title_full | Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title_fullStr | Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title_full_unstemmed | Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title_short | Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
title_sort | metabolic engineering of synechococcus elongatus pcc 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5702090/ https://www.ncbi.nlm.nih.gov/pubmed/29178875 http://dx.doi.org/10.1186/s12934-017-0824-4 |
work_keys_str_mv | AT hirokawayasutaka metabolicengineeringofsynechococcuselongatuspcc7942forimprovementof13propanediolandglycerolproductionbasedoninsilicosimulationofmetabolicfluxdistribution AT matsuoshingo metabolicengineeringofsynechococcuselongatuspcc7942forimprovementof13propanediolandglycerolproductionbasedoninsilicosimulationofmetabolicfluxdistribution AT hamadahiroyuki metabolicengineeringofsynechococcuselongatuspcc7942forimprovementof13propanediolandglycerolproductionbasedoninsilicosimulationofmetabolicfluxdistribution AT matsudafumio metabolicengineeringofsynechococcuselongatuspcc7942forimprovementof13propanediolandglycerolproductionbasedoninsilicosimulationofmetabolicfluxdistribution AT hanaitaizo metabolicengineeringofsynechococcuselongatuspcc7942forimprovementof13propanediolandglycerolproductionbasedoninsilicosimulationofmetabolicfluxdistribution |