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Title: Enhanced limonene production in cyanobacteria reveals photosynthesis limitations

Abstract

Terpenes are the major secondary metabolites produced by plants, and have diverse industrial applications as pharmaceuticals, fragrance, solvents, and biofuels. Cyanobacteria are equipped with efficient carbon fixation mechanism, and are ideal cell factories to produce various fuel and chemical products. Past efforts to produce terpenes in photosynthetic organisms have gained only limited success. Here we engineered the cyanobacterium Synechococcus elongatus PCC 7942 to efficiently produce limonene through modeling guided study. Computational modeling of limonene flux in response to photosynthetic output has revealed the downstream terpene synthase as a key metabolic flux-controlling node in the MEP (2- C-methyl-d-erythritol 4-phosphate) pathway-derived terpene biosynthesis. By enhancing the downstream limonene carbon sink, we achieved over 100-fold increase in limonene productivity, in contrast to the marginal increase achieved through stepwise metabolic engineering. The establishment of a strong limonene flux revealed potential synergy between photosynthate output and terpene biosynthesis, leading to enhanced carbon flux into the MEP pathway. Moreover, we show that enhanced limonene flux would lead to NADPH accumulation, and slow down photosynthesis electron flow. In conclusion, fine-tuning ATP/NADPH toward terpene biosynthesis could be a key parameter to adapt photosynthesis to support biofuel/bioproduct production in cyanobacteria.

Authors:
 [1];  [2];  [3];  [4];  [1];  [1];  [1];  [3];  [1];  [1];  [1]
  1. Texas A & M Univ., College Station, TX (United States)
  2. Texas A & M Univ., College Station, TX (United States); Tonghua Normal Univ., Jilin (People's Republic of China)
  3. Chinese Academy of Sciences, Shanghai (People's Republic of China)
  4. Texas A & M Univ., College Station, TX (United States); Fujian Agriculture and Forestry Univ., Fujian (People's Republic of China)
Publication Date:
Research Org.:
Texas Agrilife Research, College Station, TX (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1465396
Grant/Contract Number:  
AR0000203
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 113; Journal Issue: 50; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; photosynthesis; limonene; advanced biofuel; terpene; MEP

Citation Formats

Wang, Xin, Liu, Wei, Xin, Changpeng, Zheng, Yi, Cheng, Yanbing, Sun, Su, Li, Runze, Zhu, Xin -Guang, Dai, Susie Y., Rentzepis, Peter M., and Yuan, Joshua S.. Enhanced limonene production in cyanobacteria reveals photosynthesis limitations. United States: N. p., 2016. Web. doi:10.1073/pnas.1613340113.
Wang, Xin, Liu, Wei, Xin, Changpeng, Zheng, Yi, Cheng, Yanbing, Sun, Su, Li, Runze, Zhu, Xin -Guang, Dai, Susie Y., Rentzepis, Peter M., & Yuan, Joshua S.. Enhanced limonene production in cyanobacteria reveals photosynthesis limitations. United States. doi:10.1073/pnas.1613340113.
Wang, Xin, Liu, Wei, Xin, Changpeng, Zheng, Yi, Cheng, Yanbing, Sun, Su, Li, Runze, Zhu, Xin -Guang, Dai, Susie Y., Rentzepis, Peter M., and Yuan, Joshua S.. Wed . "Enhanced limonene production in cyanobacteria reveals photosynthesis limitations". United States. doi:10.1073/pnas.1613340113. https://www.osti.gov/servlets/purl/1465396.
@article{osti_1465396,
title = {Enhanced limonene production in cyanobacteria reveals photosynthesis limitations},
author = {Wang, Xin and Liu, Wei and Xin, Changpeng and Zheng, Yi and Cheng, Yanbing and Sun, Su and Li, Runze and Zhu, Xin -Guang and Dai, Susie Y. and Rentzepis, Peter M. and Yuan, Joshua S.},
abstractNote = {Terpenes are the major secondary metabolites produced by plants, and have diverse industrial applications as pharmaceuticals, fragrance, solvents, and biofuels. Cyanobacteria are equipped with efficient carbon fixation mechanism, and are ideal cell factories to produce various fuel and chemical products. Past efforts to produce terpenes in photosynthetic organisms have gained only limited success. Here we engineered the cyanobacterium Synechococcus elongatus PCC 7942 to efficiently produce limonene through modeling guided study. Computational modeling of limonene flux in response to photosynthetic output has revealed the downstream terpene synthase as a key metabolic flux-controlling node in the MEP (2-C-methyl-d-erythritol 4-phosphate) pathway-derived terpene biosynthesis. By enhancing the downstream limonene carbon sink, we achieved over 100-fold increase in limonene productivity, in contrast to the marginal increase achieved through stepwise metabolic engineering. The establishment of a strong limonene flux revealed potential synergy between photosynthate output and terpene biosynthesis, leading to enhanced carbon flux into the MEP pathway. Moreover, we show that enhanced limonene flux would lead to NADPH accumulation, and slow down photosynthesis electron flow. In conclusion, fine-tuning ATP/NADPH toward terpene biosynthesis could be a key parameter to adapt photosynthesis to support biofuel/bioproduct production in cyanobacteria.},
doi = {10.1073/pnas.1613340113},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 50,
volume = 113,
place = {United States},
year = {2016},
month = {11}
}

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Works referenced in this record:

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