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Title: Photosynthetic conversion of CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria

Abstract

Background: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO2 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 CO2. 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/OD730). 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 CO2 in cyanobacteria and production of squalene in S. elongatus PCCmore » 7942. Our optimized modular Over MEP 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 CO2 as bio-solar cell factories.« less

Authors:
 [1];  [2];  [2];  [3];  [4];  [2];  [2];  [5];  [6]; ORCiD logo [7]
  1. Korea Inst. of Science and Technology, Seoul (Korea, Republic of). Clean Energy Research Center; Korea Univ., Seoul (Korea, Republic of). Green School. Graduate school of Energy and Environment
  2. Korea Inst. of Science and Technology, Seoul (Korea, Republic of). Clean Energy Research Center
  3. Korea Inst. of Science and Technology, Seoul (Korea, Republic of). Clean Energy Research Center; Korea Univ., Seoul (Korea, Republic of). Dept. of Chemistry
  4. Korea Univ., Seoul (Korea, Republic of). Green School. Graduate school of Energy and Environment; Korea Univ., Seoul (Korea, Republic of). Dept. of Chemical and Biological Engineering
  5. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division
  6. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering; Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engineering
  7. Sungkyunkwan Univ., Suwon (Republic of Korea). Dept. of Food Science and Biotechnology
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1626977
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Biotechnology & Applied Microbiology; Energy & Fuels; Metabolic engineering; Cyanobacteria; Synthetic biology; Isoprenoids

Citation Formats

Choi, Sun Young, Lee, Hyun Jeong, Choi, Jaeyeon, Kim, Jiye, Sim, Sang Jun, Um, Youngsoon, Kim, Yunje, Lee, Taek Soon, Keasling, Jay D., and Woo, Han Min. Photosynthetic conversion of CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria. United States: N. p., 2016. Web. doi:10.1186/s13068-016-0617-8.
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 CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria. United States. doi:10.1186/s13068-016-0617-8.
Choi, Sun Young, Lee, Hyun Jeong, Choi, Jaeyeon, Kim, Jiye, Sim, Sang Jun, Um, Youngsoon, Kim, Yunje, Lee, Taek Soon, Keasling, Jay D., and Woo, Han Min. Thu . "Photosynthetic conversion of CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria". United States. doi:10.1186/s13068-016-0617-8. https://www.osti.gov/servlets/purl/1626977.
@article{osti_1626977,
title = {Photosynthetic conversion of CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria},
author = {Choi, Sun Young and Lee, Hyun Jeong and Choi, Jaeyeon and Kim, Jiye and Sim, Sang Jun and Um, Youngsoon and Kim, Yunje and Lee, Taek Soon and Keasling, Jay D. and Woo, Han Min},
abstractNote = {Background: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO2 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 CO2. 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/OD730). 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 CO2 in cyanobacteria and production of squalene in S. elongatus PCC 7942. Our optimized modular Over MEP 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 CO2 as bio-solar cell factories.},
doi = {10.1186/s13068-016-0617-8},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 9,
place = {United States},
year = {2016},
month = {9}
}

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