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Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus

BACKGROUND: Among the three model cyanobacterial species that have been used for engineering a system for photosynthetic production of free fatty acids (FFAs), Synechococcus elongatus PCC7942 has been the least successful; the FFA-excreting mutants constructed from this strain could attain lower rat...

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Autores principales: Kato, Akihiro, Use, Kazuhide, Takatani, Nobuyuki, Ikeda, Kazutaka, Matsuura, Miyuki, Kojima, Kouji, Aichi, Makiko, Maeda, Shin-ichi, Omata, Tatsuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4841976/
https://www.ncbi.nlm.nih.gov/pubmed/27110287
http://dx.doi.org/10.1186/s13068-016-0506-1
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author Kato, Akihiro
Use, Kazuhide
Takatani, Nobuyuki
Ikeda, Kazutaka
Matsuura, Miyuki
Kojima, Kouji
Aichi, Makiko
Maeda, Shin-ichi
Omata, Tatsuo
author_facet Kato, Akihiro
Use, Kazuhide
Takatani, Nobuyuki
Ikeda, Kazutaka
Matsuura, Miyuki
Kojima, Kouji
Aichi, Makiko
Maeda, Shin-ichi
Omata, Tatsuo
author_sort Kato, Akihiro
collection PubMed
description BACKGROUND: Among the three model cyanobacterial species that have been used for engineering a system for photosynthetic production of free fatty acids (FFAs), Synechococcus elongatus PCC7942 has been the least successful; the FFA-excreting mutants constructed from this strain could attain lower rates of FFA excretion and lower final FFA concentrations than the mutants constructed from Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002. It has been suggested that S. elongatus PCC7942 cells suffer from toxicity of FFA, but the cause of the low productivity has remained to be determined. RESULTS: By modulating the expression level of the acyl–acyl carrier protein thioesterase and raising the light intensity during cultivation, FFA secretion rates comparable to those obtained with the other cyanobacterial species were attained with an engineered Synechococcus elongatus mutant (dAS1T). The final FFA concentration in the external medium was also higher than previously reported for other S. elongatus mutants. However, about 85 % of the total FFA in the culture was found to remain in the cells, causing severe photoinhibition. Targeted inactivation of the wzt gene in dAS1T, which gene manipulation was previously shown to result in loss of the hydrophilic O-antigen layer on the cell surface, increased FFA secretion, alleviated photoinhibition, and lead to 50 and 45 % increase in the final cell density and the total amount of FFA in the culture (i.e., the sum of the cellular and extracellular FFA), respectively. The average rate of production of total FFA by the culture of the ∆wzt strain was 2.7 mg L(−1) h(−1), being five times higher than those reported for Synechocystis sp. PCC 6803 and comparable to the rates of triacylglycerol production in green algae. CONCLUSION: Synechococcus elongatus PCC7942 has larger capacity of FFA production than Synechocystis sp. PCC6803 but accumulates most of the product in the cell because of the imbalance of the rates of FFA production and secretion. This causes severe photoinhibition and exerts adverse effects on cell growth and FFA productivity. Enhancement of FFA secretion would be required to fully exploiting the capacity of FFA production for the purpose of biofuel production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0506-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-48419762016-04-24 Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus Kato, Akihiro Use, Kazuhide Takatani, Nobuyuki Ikeda, Kazutaka Matsuura, Miyuki Kojima, Kouji Aichi, Makiko Maeda, Shin-ichi Omata, Tatsuo Biotechnol Biofuels Research BACKGROUND: Among the three model cyanobacterial species that have been used for engineering a system for photosynthetic production of free fatty acids (FFAs), Synechococcus elongatus PCC7942 has been the least successful; the FFA-excreting mutants constructed from this strain could attain lower rates of FFA excretion and lower final FFA concentrations than the mutants constructed from Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002. It has been suggested that S. elongatus PCC7942 cells suffer from toxicity of FFA, but the cause of the low productivity has remained to be determined. RESULTS: By modulating the expression level of the acyl–acyl carrier protein thioesterase and raising the light intensity during cultivation, FFA secretion rates comparable to those obtained with the other cyanobacterial species were attained with an engineered Synechococcus elongatus mutant (dAS1T). The final FFA concentration in the external medium was also higher than previously reported for other S. elongatus mutants. However, about 85 % of the total FFA in the culture was found to remain in the cells, causing severe photoinhibition. Targeted inactivation of the wzt gene in dAS1T, which gene manipulation was previously shown to result in loss of the hydrophilic O-antigen layer on the cell surface, increased FFA secretion, alleviated photoinhibition, and lead to 50 and 45 % increase in the final cell density and the total amount of FFA in the culture (i.e., the sum of the cellular and extracellular FFA), respectively. The average rate of production of total FFA by the culture of the ∆wzt strain was 2.7 mg L(−1) h(−1), being five times higher than those reported for Synechocystis sp. PCC 6803 and comparable to the rates of triacylglycerol production in green algae. CONCLUSION: Synechococcus elongatus PCC7942 has larger capacity of FFA production than Synechocystis sp. PCC6803 but accumulates most of the product in the cell because of the imbalance of the rates of FFA production and secretion. This causes severe photoinhibition and exerts adverse effects on cell growth and FFA productivity. Enhancement of FFA secretion would be required to fully exploiting the capacity of FFA production for the purpose of biofuel production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0506-1) contains supplementary material, which is available to authorized users. BioMed Central 2016-04-23 /pmc/articles/PMC4841976/ /pubmed/27110287 http://dx.doi.org/10.1186/s13068-016-0506-1 Text en © Kato et al. 2016 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
Kato, Akihiro
Use, Kazuhide
Takatani, Nobuyuki
Ikeda, Kazutaka
Matsuura, Miyuki
Kojima, Kouji
Aichi, Makiko
Maeda, Shin-ichi
Omata, Tatsuo
Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title_full Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title_fullStr Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title_full_unstemmed Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title_short Modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium Synechococcus elongatus
title_sort modulation of the balance of fatty acid production and secretion is crucial for enhancement of growth and productivity of the engineered mutant of the cyanobacterium synechococcus elongatus
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4841976/
https://www.ncbi.nlm.nih.gov/pubmed/27110287
http://dx.doi.org/10.1186/s13068-016-0506-1
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