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Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria

BACKGROUND: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO(2) to value-added chemicals as bio-solar cell factories. However, the production levels of isoprenoids in engineered cyanobacteria were quite low, compared to other microbial hosts. Therefore, modular opti...

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Autores principales: Choi, Sun Young, Lee, Hyun Jeong, Choi, Jaeyeon, Kim, Jiye, Sim, Sang Jun, Um, Youngsoon, Kim, Yunje, Lee, Taek Soon, Keasling, Jay D., Woo, Han Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034544/
https://www.ncbi.nlm.nih.gov/pubmed/27688805
http://dx.doi.org/10.1186/s13068-016-0617-8
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author Choi, Sun Young
Lee, Hyun Jeong
Choi, Jaeyeon
Kim, Jiye
Sim, Sang Jun
Um, Youngsoon
Kim, Yunje
Lee, Taek Soon
Keasling, Jay D.
Woo, Han Min
author_facet Choi, Sun Young
Lee, Hyun Jeong
Choi, Jaeyeon
Kim, Jiye
Sim, Sang Jun
Um, Youngsoon
Kim, Yunje
Lee, Taek Soon
Keasling, Jay D.
Woo, Han Min
author_sort Choi, Sun Young
collection PubMed
description BACKGROUND: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO(2) to value-added chemicals as bio-solar cell factories. However, the production levels of isoprenoids in engineered cyanobacteria were quite low, compared to other microbial hosts. Therefore, modular optimization of multiple gene expressions for metabolic engineering of cyanobacteria is required for the production of farnesyl diphosphate-derived isoprenoids from CO(2). RESULTS: Here, we engineered Synechococcus elongatus PCC 7942 with modular metabolic pathways consisting of the methylerythritol phosphate pathway enzymes and the amorphadiene synthase for production of amorpha-4,11-diene, resulting in significantly increased levels (23-fold) of amorpha-4,11-diene (19.8 mg/L) in the best strain relative to a parental strain. Replacing amorphadiene synthase with squalene synthase led to the synthesis of a high amount of squalene (4.98 mg/L/OD(730)). Overexpression of farnesyl diphosphate synthase is the most critical factor for the significant production, whereas overexpression of 1-deoxy-d-xylulose 5-phosphate reductase is detrimental to the cell growth and the production. Additionally, the cyanobacterial growth inhibition was alleviated by expressing a terpene synthase in S. elongatus PCC 7942 strain with the optimized MEP pathway only (SeHL33). CONCLUSIONS: This is the first demonstration of photosynthetic production of amorpha-4,11-diene from CO(2) in cyanobacteria and production of squalene in S. elongatus PCC 7942. Our optimized modular OverMEP strain (SeHL33) with either co-expression of ADS or SQS demonstrated the highest production levels of amorpha-4,11-diene and squalene, which could expand the list of farnesyl diphosphate-derived isoprenoids from CO(2) as bio-solar cell factories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0617-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-50345442016-09-29 Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria Choi, Sun Young Lee, Hyun Jeong Choi, Jaeyeon Kim, Jiye Sim, Sang Jun Um, Youngsoon Kim, Yunje Lee, Taek Soon Keasling, Jay D. Woo, Han Min Biotechnol Biofuels Research BACKGROUND: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO(2) to value-added chemicals as bio-solar cell factories. However, the production levels of isoprenoids in engineered cyanobacteria were quite low, compared to other microbial hosts. Therefore, modular optimization of multiple gene expressions for metabolic engineering of cyanobacteria is required for the production of farnesyl diphosphate-derived isoprenoids from CO(2). RESULTS: Here, we engineered Synechococcus elongatus PCC 7942 with modular metabolic pathways consisting of the methylerythritol phosphate pathway enzymes and the amorphadiene synthase for production of amorpha-4,11-diene, resulting in significantly increased levels (23-fold) of amorpha-4,11-diene (19.8 mg/L) in the best strain relative to a parental strain. Replacing amorphadiene synthase with squalene synthase led to the synthesis of a high amount of squalene (4.98 mg/L/OD(730)). Overexpression of farnesyl diphosphate synthase is the most critical factor for the significant production, whereas overexpression of 1-deoxy-d-xylulose 5-phosphate reductase is detrimental to the cell growth and the production. Additionally, the cyanobacterial growth inhibition was alleviated by expressing a terpene synthase in S. elongatus PCC 7942 strain with the optimized MEP pathway only (SeHL33). CONCLUSIONS: This is the first demonstration of photosynthetic production of amorpha-4,11-diene from CO(2) in cyanobacteria and production of squalene in S. elongatus PCC 7942. Our optimized modular OverMEP strain (SeHL33) with either co-expression of ADS or SQS demonstrated the highest production levels of amorpha-4,11-diene and squalene, which could expand the list of farnesyl diphosphate-derived isoprenoids from CO(2) as bio-solar cell factories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0617-8) contains supplementary material, which is available to authorized users. BioMed Central 2016-09-22 /pmc/articles/PMC5034544/ /pubmed/27688805 http://dx.doi.org/10.1186/s13068-016-0617-8 Text en © The Author(s) 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
Choi, Sun Young
Lee, Hyun Jeong
Choi, Jaeyeon
Kim, Jiye
Sim, Sang Jun
Um, Youngsoon
Kim, Yunje
Lee, Taek Soon
Keasling, Jay D.
Woo, Han Min
Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title_full Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title_fullStr Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title_full_unstemmed Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title_short Photosynthetic conversion of CO(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
title_sort photosynthetic conversion of co(2) to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034544/
https://www.ncbi.nlm.nih.gov/pubmed/27688805
http://dx.doi.org/10.1186/s13068-016-0617-8
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