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Title: Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet

Abstract

Traditional bioproduct engineering focuses on pathway optimization, yet is often complicated by product inhibition, downstream consumption, and the toxicity of certain products. Here, we present the co-compartmentation of biosynthesis and storage via a synthetic droplet as an effective new strategy to improve the bioproduct yield, with squalene as a model compound. A hydrophobic protein was designed and introduced into the tobacco chloroplast to generate a synthetic droplet for terpene storage. Simultaneously, squalene biosynthesis enzymes were introduced to chloroplasts together with the droplet-forming protein to co-compartmentalize the biosynthesis and storage of squalene. The strategy has enabled a record yield of squalene at 2.6 mg/g fresh weight without compromising plant growth. Confocal fluorescent microscopy imaging, stimulated Raman scattering microscopy, and droplet composition analysis confirmed the formation of synthetic storage droplet in chloroplast. The co-compartmentation of synthetic storage droplet with a targeted metabolic pathway engineering represents a new strategy for enhancing bioproduct yield.

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1];  [3]; ORCiD logo [4]; ORCiD logo [1]
  1. Texas A & M Univ., College Station, TX (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  3. Univ. of Iowa, Coralville, IA (United States). State Hygenic Lab.
  4. Michigan State Univ., East Lansing, MI (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1424893
Report Number(s):
NREL/JA-2700-71061
Journal ID: ISSN 2161-5063
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
ACS Synthetic Biology
Additional Journal Information:
Journal Volume: 7; Journal Issue: 3; Journal ID: ISSN 2161-5063
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; co-compartmentation; hydrophobic protein; oleosin; squalene; synthetic droplet

Citation Formats

Zhao, Cheng, Kim, YongKyoung, Zeng, Yining, Li, Man, Wang, Xin, Hu, Cheng, Gorman, Connor, Dai, Susie Y., Ding, Shi-You, and Yuan, Joshua S. Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet. United States: N. p., 2018. Web. doi:10.1021/acssynbio.7b00368.
Zhao, Cheng, Kim, YongKyoung, Zeng, Yining, Li, Man, Wang, Xin, Hu, Cheng, Gorman, Connor, Dai, Susie Y., Ding, Shi-You, & Yuan, Joshua S. Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet. United States. doi:10.1021/acssynbio.7b00368.
Zhao, Cheng, Kim, YongKyoung, Zeng, Yining, Li, Man, Wang, Xin, Hu, Cheng, Gorman, Connor, Dai, Susie Y., Ding, Shi-You, and Yuan, Joshua S. Tue . "Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet". United States. doi:10.1021/acssynbio.7b00368. https://www.osti.gov/servlets/purl/1424893.
@article{osti_1424893,
title = {Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet},
author = {Zhao, Cheng and Kim, YongKyoung and Zeng, Yining and Li, Man and Wang, Xin and Hu, Cheng and Gorman, Connor and Dai, Susie Y. and Ding, Shi-You and Yuan, Joshua S.},
abstractNote = {Traditional bioproduct engineering focuses on pathway optimization, yet is often complicated by product inhibition, downstream consumption, and the toxicity of certain products. Here, we present the co-compartmentation of biosynthesis and storage via a synthetic droplet as an effective new strategy to improve the bioproduct yield, with squalene as a model compound. A hydrophobic protein was designed and introduced into the tobacco chloroplast to generate a synthetic droplet for terpene storage. Simultaneously, squalene biosynthesis enzymes were introduced to chloroplasts together with the droplet-forming protein to co-compartmentalize the biosynthesis and storage of squalene. The strategy has enabled a record yield of squalene at 2.6 mg/g fresh weight without compromising plant growth. Confocal fluorescent microscopy imaging, stimulated Raman scattering microscopy, and droplet composition analysis confirmed the formation of synthetic storage droplet in chloroplast. The co-compartmentation of synthetic storage droplet with a targeted metabolic pathway engineering represents a new strategy for enhancing bioproduct yield.},
doi = {10.1021/acssynbio.7b00368},
journal = {ACS Synthetic Biology},
issn = {2161-5063},
number = 3,
volume = 7,
place = {United States},
year = {2018},
month = {2}
}

Journal Article:
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