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:
-
[4];
[1]
- Texas A & M Univ., College Station, TX (United States)
- National Renewable Energy Lab. (NREL), Golden, CO (United States)
- Univ. of Iowa, Coralville, IA (United States). State Hygenic Lab.
- Michigan State Univ., East Lansing, MI (United States)
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (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:
- 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. https://doi.org/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. https://doi.org/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},
number = 3,
volume = 7,
place = {United States},
year = {Tue Feb 13 00:00:00 EST 2018},
month = {Tue Feb 13 00:00:00 EST 2018}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 24 works
Citation information provided by
Web of Science
Web of Science
Figures / Tables:
Figure 1: Treatment experiments in darkness and the design of synthetic droplet. (A) Squalene accumulation of FPS-SQS lines in leaves for 15 days in the condition of 16 h light/8 h dark with triplicates. (B) Squalene accumulation of FPS-SQS lines in leaves for 15 days of the dark treatment withmore »
All figures and tables
(4 total)
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Manufacturing Molecules Through Metabolic Engineering
journal, December 2010
- Keasling, Jay D.
- Science, Vol. 330, Issue 6009, p. 1355-1358
Squalene: A natural triterpene for use in disease management and therapy
journal, December 2009
- Reddy, L. Harivardhan; Couvreur, Patrick
- Advanced Drug Delivery Reviews, Vol. 61, Issue 15
Photosynthetic terpene hydrocarbon production for fuels and chemicals
journal, January 2015
- Wang, Xin; Ort, Donald R.; Yuan, Joshua S.
- Plant Biotechnology Journal, Vol. 13, Issue 2
Production of Squalene in Synechocystis sp. PCC 6803
journal, March 2014
- Englund, Elias; Pattanaik, Bagmi; Ubhayasekera, Sarojini Jayantha K.
- PLoS ONE, Vol. 9, Issue 3
Arabidopsis thaliana Squalene Epoxidase 1 Is Essential for Root and Seed Development
journal, April 2007
- Rasbery, Jeanne M.; Shan, Hui; LeClair, Renee J.
- Journal of Biological Chemistry, Vol. 282, Issue 23
Methylerythritol 4-phosphate (MEP) pathway metabolic regulation
journal, January 2014
- Banerjee, A.; Sharkey, T. D.
- Nat. Prod. Rep., Vol. 31, Issue 8
Network Analysis of the MVA and MEP Pathways for Isoprenoid Synthesis
journal, April 2013
- Vranová, Eva; Coman, Diana; Gruissem, Wilhelm
- Annual Review of Plant Biology, Vol. 64, Issue 1
Alleviating monoterpene toxicity using a two-phase extractive fermentation for the bioproduction of jet fuel mixtures in Saccharomyces cerevisiae
journal, May 2012
- Brennan, Timothy C. R.; Turner, Christopher D.; Krömer, Jens O.
- Biotechnology and Bioengineering, Vol. 109, Issue 10
Transcriptome analysis of alfalfa glandular trichomes
journal, December 2004
- Aziz, Naveed; Paiva, Nancy L.; May, Gregory D.
- Planta, Vol. 221, Issue 1
Divergent Regulation of Terpenoid Metabolism in the Trichomes of Wild and Cultivated Tomato Species
journal, November 2008
- Besser, Katrin; Harper, Andrea; Welsby, Nicholas
- Plant Physiology, Vol. 149, Issue 1
Terpenoid Biosynthesis and Specialized Vascular Cells of Conifer Defense
journal, January 2010
- Zulak, Katherine G.; Bohlmann, Jörg
- Journal of Integrative Plant Biology, Vol. 52, Issue 1
Engineering triterpene metabolism in tobacco
journal, June 2012
- Wu, Shuiqin; Jiang, Zuodong; Kempinski, Chase
- Planta, Vol. 236, Issue 3
Linear Relationships between Lipophilic Character and Biological Activity of Drugs
journal, January 1972
- Hansch, Corwin; Dunn, William J.
- Journal of Pharmaceutical Sciences, Vol. 61, Issue 1
Membrane topology and sequence requirements for oil body targeting of oleosin: Role of oleosin topology in oil body targeting
journal, February 2004
- Abell, Ben M.; Hahn, Margaret; Holbrook, Larry A.
- The Plant Journal, Vol. 37, Issue 4
Identification of a New Class of Lipid Droplet-Associated Proteins in Plants
journal, July 2013
- Horn, Patrick J.; James, Christopher N.; Gidda, Satinder K.
- Plant Physiology, Vol. 162, Issue 4
In Vivo Packaging of Triacylglycerols Enhances Arabidopsis Leaf Biomass and Energy Density
journal, April 2013
- Winichayakul, Somrutai; Scott, Richard William; Roldan, Marissa
- Plant Physiology, Vol. 162, Issue 2
Lipid Droplets and Cellular Lipid Metabolism
journal, July 2012
- Walther, Tobias C.; Farese, Robert V.
- Annual Review of Biochemistry, Vol. 81, Issue 1
Role of the Proline Knot Motif in Oleosin Endoplasmic Reticulum Topology and Oil Body Targeting
journal, August 1997
- Abell, Ben M.; Holbrook, Larry A.; Abenes, Malleva
- The Plant Cell, Vol. 9, Issue 8
Membrane Protein Topology of Oleosin Is Constrained by Its Long Hydrophobic Domain
journal, October 2001
- Abell, Ben M.; High, Stephen; Moloney, Maurice M.
- Journal of Biological Chemistry, Vol. 277, Issue 10
Colony Organization in the Green Alga Botryococcus braunii (Race B) Is Specified by a Complex Extracellular Matrix
journal, August 2012
- Weiss, Taylor L.; Roth, Robyn; Goodson, Carrie
- Eukaryotic Cell, Vol. 11, Issue 12
Raman imaging to investigate ultrastructure and composition of plant cell walls: distribution of lignin and cellulose in black spruce wood (Picea mariana)
journal, June 2006
- Agarwal, Umesh P.
- Planta, Vol. 224, Issue 5
Chemical Imaging of Poplar Wood Cell Walls by Confocal Raman Microscopy
journal, February 2006
- Gierlinger, Notburga; Schwanninger, Manfred
- Plant Physiology, Vol. 140, Issue 4
Label-free in situ imaging of lignification in the cell wall of low lignin transgenic Populus trichocarpa
journal, June 2009
- Schmidt, M.; Schwartzberg, A. M.; Perera, P. N.
- Planta, Vol. 230, Issue 3
Understanding tissue specific compositions of bioenergy feedstocks through hyperspectral Raman imaging
journal, October 2010
- Sun, Lan; Simmons, Blake A.; Singh, Seema
- Biotechnology and Bioengineering, Vol. 108, Issue 2
Efficient coagulation of microalgae in cultures with filamentous fungi
journal, January 2013
- Xie, Shangxian; Sun, Su; Dai, Susie Y.
- Algal Research, Vol. 2, Issue 1
Engineering Triterpene and Methylated Triterpene Production in Plants Provides Biochemical and Physiological Insights into Terpene Metabolism
journal, November 2015
- Jiang, Zuodong; Kempinski, Chase; Bush, Caroline J.
- Plant Physiology, Vol. 170, Issue 2
MF59 adjuvant: the best insurance against influenza strain diversity
journal, April 2011
- O’Hagan, Derek T.; Rappuoli, Rino; De Gregorio, Ennio
- Expert Review of Vaccines, Vol. 10, Issue 4
Tobacco as a production platform for biofuel overexpression of Arabidopsis DGAT and LEC2 genes increases accumulation and shifts the composition of lipids in green biomass
journal, April 2010
- Andrianov, Vyacheslav; Borisjuk, Nikolai; Pogrebnyak, Natalia
- Plant Biotechnology Journal, Vol. 8, Issue 3, p. 277-287
Starch Changes in Developing and Senescing Tobacco Leaves
journal, January 1962
- Matheson, Nk; Wheatley, Jm
- Australian Journal of Biological Sciences, Vol. 15, Issue 3
Production of squalene by microbes: an update
journal, October 2016
- Xu, Wen; Ma, Xi; Wang, Yang
- World Journal of Microbiology and Biotechnology, Vol. 32, Issue 12
Enzymatic assembly of DNA molecules up to several hundred kilobases
journal, April 2009
- Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan
- Nature Methods, Vol. 6, Issue 5, p. 343-345
Isolation of Chloroplasts from Plant Protoplasts
journal, October 2015
- Lung, Shiu-Cheung; Smith, Matthew D.; Chuong, Simon D. X.
- Cold Spring Harbor Protocols, Vol. 2015, Issue 10
Label-Free, Real-Time Monitoring of Biomass Processing with Stimulated Raman Scattering Microscopy
journal, June 2010
- Saar, Brian G.; Zeng, Yining; Freudiger, Christian W.
- Angewandte Chemie International Edition, Vol. 49, Issue 32
Imaging Lignin-Downregulated Alfalfa Using Coherent Anti-Stokes Raman Scattering Microscopy
journal, February 2010
- Zeng, Yining; Saar, Brian G.; Friedrich, Marcel G.
- BioEnergy Research, Vol. 3, Issue 3
Lignin plays a negative role in the biochemical process for producing lignocellulosic biofuels
journal, June 2014
- Zeng, Yining; Zhao, Shuai; Yang, Shihui
- Current Opinion in Biotechnology, Vol. 27
In situ micro-spectroscopic investigation of lignin in poplar cell walls pretreated by maleic acid
journal, August 2015
- Zeng, Yining; Zhao, Shuai; Wei, Hui
- Biotechnology for Biofuels, Vol. 8, Issue 1
Works referencing / citing this record:
Mouse lipogenic proteins promote the co-accumulation of triacylglycerols and sesquiterpenes in plant cells
journal, March 2019
- Cai, Yingqi; Whitehead, Payton; Chappell, Joe
- Planta, Vol. 250, Issue 1
Cytosolic lipid droplets as engineered organelles for production and accumulation of terpenoid biomaterials in leaves
journal, February 2019
- Sadre, Radin; Kuo, Peiyen; Chen, Jiaxing
- Nature Communications, Vol. 10, Issue 1
Dynamic metabolic solutions to the sessile life style of plants
journal, January 2018
- Knudsen, Camilla; Gallage, Nethaji Janeshawari; Hansen, Cecilie Cetti
- Natural Product Reports, Vol. 35, Issue 11
Cytosolic lipid droplets as engineered organelles for production and accumulation of terpenoid biomaterials in leaves
journal, February 2019
- Sadre, Radin; Kuo, Peiyen; Chen, Jiaxing
- Nature Communications, Vol. 10, Issue 1
Dynamic metabolic solutions to the sessile life style of plants
journal, January 2018
- Knudsen, Camilla; Gallage, Nethaji Janeshawari; Hansen, Cecilie Cetti
- Natural Product Reports, Vol. 35, Issue 11
Figures / Tables found in this record:
Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.