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Title: Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil

Abstract

Modified fatty acids (mFA) have diverse uses, e.g., cyclopropane fatty acids (CPA) are feedstocks for producing coatings, lubricants, plastics, and cosmetics. The expression of mFA-producing enzymes in crop and model plants generally results in lower levels of mFA accumulation than in their natural-occurring source plants. In order to further our understanding of metabolic bottlenecks that limit mFA accumulation, we generated transgenic Camelina sativa lines co-expressing Escherichia coli cyclopropane synthase (EcCPS) and Sterculia foetida lysophosphatidic acid acyltransferase (SfLPAT). In contrast to transgenic CPA-accumulating Arabidopsis, CPA accumulation in camelina caused only minor changes in seed weight, germination rate, oil accumulation, and seedling development. CPA accumulated to much higher levels in membrane than storage lipids, comprising more than 60% of total fatty acid in both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) versus 26% in diacylglycerol (DAG) and 12% in triacylglycerol (TAG) indicating bottlenecks in the transfer of CPA from PC to DAG and from DAG to TAG. Upon coexpression of SfLPAT with EcCPS, di-CPA-PC increased by ~50% relative to lines expressing EcCPS alone with the di-CPA-PC primarily observed in the embryonic axis and mono-CPA-PC primarily in cotyledon tissue. EcCPS-SfLPAT lines revealed a redistribution of CPA from the sn-1 to sn-2 positions within PC andmore » PE that was associated with a doubling of CPA accumulation in both DAG and TAG. Finally, the identification of metabolic bottlenecks in acyl transfer between site of synthesis (phospholipids) and deposition in storage oils (TAGs) lays the foundation for the optimizing CPA accumulation through directed engineering of oil synthesis in target crops.« less

Authors:
 [1];  [2];  [3];  [4];  [1];  [5];  [4];  [3];  [2];  [6];  [6]
  1. Stony Brook Univ., NY (United States). Dept. of Chemistry. Biochemistry and Cell Biology Dept.
  2. Univ. of Nebrask. Center for Plant Science Innovation and Dept. of Biochemistry
  3. Univ. of North Texas, Denton, TX (United States). Center for Advanced Scientific Computing and Modeling (CASCaM). Dept. of Biological Sciences
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Biology Dept.
  5. Stony Brook Univ., NY (United States). Biochemistry and Cell Biology Dept.
  6. Stony Brook Univ., NY (United States). Dept. of Chemistry. Biochemistry and Cell Biology Dept.; Brookhaven National Lab. (BNL), Upton, NY (United States). Biology Dept.
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)
OSTI Identifier:
1412647
Report Number(s):
BNL-114333-2017-JA
Journal ID: ISSN 1467-7644
Grant/Contract Number:  
SC0012704; FG02-09ER64812; KC0304000; DBI 1117680
Resource Type:
Accepted Manuscript
Journal Name:
Plant Biotechnology Journal
Additional Journal Information:
Journal Volume: 16; Journal Issue: 4; Journal ID: ISSN 1467-7644
Publisher:
Society for Experimental Biology; Association of Applied Biology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; cyclopropane fatty acid; Camelina sativa; unusual fatty acid; lipid metabolism; triacylglycerol; lipid synthesis

Citation Formats

Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, and Shanklin, John. Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil. United States: N. p., 2017. Web. doi:10.1111/pbi.12839.
Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, & Shanklin, John. Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil. United States. https://doi.org/10.1111/pbi.12839
Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, and Shanklin, John. Wed . "Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil". United States. https://doi.org/10.1111/pbi.12839. https://www.osti.gov/servlets/purl/1412647.
@article{osti_1412647,
title = {Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil},
author = {Yu, Xiao-Hong and Cahoon, Rebecca E. and Horn, Patrick J. and Shi, Hai and Prakash, Richa Rawat and Cai, Yuanheng and Hearney, Maegan and Chapman, Kent D. and Cahoon, Edgar B. and Schwender, Jorg and Shanklin, John},
abstractNote = {Modified fatty acids (mFA) have diverse uses, e.g., cyclopropane fatty acids (CPA) are feedstocks for producing coatings, lubricants, plastics, and cosmetics. The expression of mFA-producing enzymes in crop and model plants generally results in lower levels of mFA accumulation than in their natural-occurring source plants. In order to further our understanding of metabolic bottlenecks that limit mFA accumulation, we generated transgenic Camelina sativa lines co-expressing Escherichia coli cyclopropane synthase (EcCPS) and Sterculia foetida lysophosphatidic acid acyltransferase (SfLPAT). In contrast to transgenic CPA-accumulating Arabidopsis, CPA accumulation in camelina caused only minor changes in seed weight, germination rate, oil accumulation, and seedling development. CPA accumulated to much higher levels in membrane than storage lipids, comprising more than 60% of total fatty acid in both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) versus 26% in diacylglycerol (DAG) and 12% in triacylglycerol (TAG) indicating bottlenecks in the transfer of CPA from PC to DAG and from DAG to TAG. Upon coexpression of SfLPAT with EcCPS, di-CPA-PC increased by ~50% relative to lines expressing EcCPS alone with the di-CPA-PC primarily observed in the embryonic axis and mono-CPA-PC primarily in cotyledon tissue. EcCPS-SfLPAT lines revealed a redistribution of CPA from the sn-1 to sn-2 positions within PC and PE that was associated with a doubling of CPA accumulation in both DAG and TAG. Finally, the identification of metabolic bottlenecks in acyl transfer between site of synthesis (phospholipids) and deposition in storage oils (TAGs) lays the foundation for the optimizing CPA accumulation through directed engineering of oil synthesis in target crops.},
doi = {10.1111/pbi.12839},
journal = {Plant Biotechnology Journal},
number = 4,
volume = 16,
place = {United States},
year = {2017},
month = {9}
}

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Works referenced in this record:

The Significance of Different Diacylgycerol Synthesis Pathways on Plant Oil Composition and Bioengineering
journal, January 2012


Biology and Biochemistry of Plant Phospholipases
journal, May 2011

  • Chen, Guanqun; Snyder, Crystal L.; Greer, Michael S.
  • Critical Reviews in Plant Sciences, Vol. 30, Issue 3
  • DOI: 10.1080/07352689.2011.572033

Redirection of metabolic flux for high levels of omega-7 monounsaturated fatty acid accumulation in camelina seeds
journal, July 2014

  • Nguyen, Huu Tam; Park, Hyunwoo; Koster, Karen L.
  • Plant Biotechnology Journal, Vol. 13, Issue 1
  • DOI: 10.1111/pbi.12233

Carbocyclic fatty acids in plants: Biochemical and molecular genetic characterization of cyclopropane fatty acid synthesis of Sterculia foetida
journal, May 2002

  • Bao, X.; Katz, S.; Pollard, M.
  • Proceedings of the National Academy of Sciences, Vol. 99, Issue 10
  • DOI: 10.1073/pnas.092152999

Synthetic redesign of plant lipid metabolism
journal, June 2016

  • Haslam, Richard P.; Sayanova, Olga; Kim, Hae Jin
  • The Plant Journal, Vol. 87, Issue 1
  • DOI: 10.1111/tpj.13172

Modulating seed β-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil
journal, March 2007

  • Pidkowich, M. S.; Nguyen, H. T.; Heilmann, I.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 11, p. 4742-4747
  • DOI: 10.1073/pnas.0611141104

The emerging biofuel crop Camelina sativa retains a highly undifferentiated hexaploid genome structure
journal, April 2014

  • Kagale, Sateesh; Koh, Chushin; Nixon, John
  • Nature Communications, Vol. 5, Issue 1
  • DOI: 10.1038/ncomms4706

Fatty acid synthesis is inhibited by inefficient utilization of unusual fatty acids for glycerolipid assembly
journal, January 2014

  • Bates, P. D.; Johnson, S. R.; Cao, X.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 3
  • DOI: 10.1073/pnas.1318511111

Conjugated fatty acids accumulate to high levels in phospholipids of metabolically engineered soybean and Arabidopsis seeds
journal, June 2006


WRINKLED1 Rescues Feedback Inhibition of Fatty Acid Synthesis in Hydroxylase-Expressing Seeds
journal, March 2016

  • Adhikari, Neil D.; Bates, Philip D.; Browse, John
  • Plant Physiology, Vol. 171, Issue 1
  • DOI: 10.1104/pp.15.01906

Fatty Acid Distribution in Bacterial Phospholipids. The Specificity of the Cyclopropane Synthetase Reaction *
journal, September 1964


Metabolic Interactions between the Lands Cycle and the Kennedy Pathway of Glycerolipid Synthesis in Arabidopsis Developing Seeds
journal, November 2012

  • Wang, Liping; Shen, Wenyun; Kazachkov, Michael
  • The Plant Cell, Vol. 24, Issue 11
  • DOI: 10.1105/tpc.112.104604

Characterization of Oilseed Lipids from “DHA-Producing Camelina sativa”: A New Transformed Land Plant Containing Long-Chain Omega-3 Oils
journal, February 2014

  • Mansour, Maged; Shrestha, Pushkar; Belide, Srinivas
  • Nutrients, Vol. 6, Issue 2
  • DOI: 10.3390/nu6020776

Camelina seed transcriptome: a tool for meal and oil improvement and translational research
journal, April 2013

  • Nguyen, Huu T.; Silva, Jillian E.; Podicheti, Ram
  • Plant Biotechnology Journal, Vol. 11, Issue 6
  • DOI: 10.1111/pbi.12068

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil
journal, February 2017

  • Marmon, Sofia; Sturtevant, Drew; Herrfurth, Cornelia
  • Plant Physiology, Vol. 173, Issue 4
  • DOI: 10.1104/pp.16.01865

Litchi chinensis fatty acid diversity: occurrence of the unusual cyclopropanoic fatty acids
journal, December 2000

  • Gontier, E.; Boussouel, N.; Terrasse, C.
  • Biochemical Society Transactions, Vol. 28, Issue 6
  • DOI: 10.1042/bst0280578

Comparative Transcriptomic Analysis of Developing Cotton Cotyledons and Embryo Axis
journal, August 2013


Phospholipase Dζ Enhances Diacylglycerol Flux into Triacylglycerol
journal, March 2017

  • Yang, Wenyu; Wang, Geliang; Li, Jia
  • Plant Physiology, Vol. 174, Issue 1
  • DOI: 10.1104/pp.17.00026

Metabolic engineering of hydroxy fatty acid production in plants: RcDGAT2 drives dramatic increases in ricinoleate levels in seed oil
journal, October 2008


Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation
journal, September 2007


Cyclopropane ring formation in membrane lipids of bacteria.
journal, January 1997

  • Grogan, D. W.; Cronan, J. E.
  • Microbiology and molecular biology reviews : MMBR, Vol. 61, Issue 4
  • DOI: 10.1128/.61.4.429-441.1997

Desaturation and Hydroxylation: RESIDUES 148 AND 324 OF
journal, February 2002

  • Broadwater, John A.; Whittle, Edward; Shanklin, John
  • Journal of Biological Chemistry, Vol. 277, Issue 18
  • DOI: 10.1074/jbc.M200231200

Vernonia DGATs can complement the disrupted oil and protein metabolism in epoxygenase-expressing soybean seeds
journal, January 2012


Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis.
journal, January 1994

  • Okuley, J.; Lightner, J.; Feldmann, K.
  • The Plant Cell, Vol. 6, Issue 1
  • DOI: 10.1105/tpc.6.1.147

Characterization and analysis of the cotton cyclopropane fatty acid synthase family and their contribution to cyclopropane fatty acid synthesis
journal, January 2011


Characterization of Cyclopropane Fatty-acid Synthase from Sterculia foetida
journal, January 2003

  • Bao, Xiaoming; Thelen, Jay J.; Bonaventure, Gustavo
  • Journal of Biological Chemistry, Vol. 278, Issue 15
  • DOI: 10.1074/jbc.M212464200

Understanding the control of acyl flux through the lipid metabolic network of plant oil biosynthesis
journal, September 2016

  • Bates, Philip D.
  • Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1861, Issue 9
  • DOI: 10.1016/j.bbalip.2016.03.021

The plant lipidome in human and environmental health
journal, September 2016


Castor Phospholipid:Diacylglycerol Acyltransferase Facilitates Efficient Metabolism of Hydroxy Fatty Acids in Transgenic Arabidopsis
journal, December 2010

  • van Erp, Harrie; Bates, Philip D.; Burgal, Julie
  • Plant Physiology, Vol. 155, Issue 2
  • DOI: 10.1104/pp.110.167239

Replacing fossil oil with fresh oil - with what and for what?
journal, April 2011

  • Carlsson, Anders S.; Yilmaz, Jenny Lindberg; Green, Allan G.
  • European Journal of Lipid Science and Technology, Vol. 113, Issue 7
  • DOI: 10.1002/ejlt.201100032

Trialkylsulfonium- and Trialkylselenoniumhydroxides for the Pyrolytic Alkylation of Acidic Compounds
journal, January 1982


Metabolite Imager: customized spatial analysis of metabolite distributions in mass spectrometry imaging
journal, August 2013


An enzyme regulating triacylglycerol composition is encoded by the ROD1 gene of Arabidopsis
journal, October 2009

  • Lu, C.; Xin, Z.; Ren, Z.
  • Proceedings of the National Academy of Sciences, Vol. 106, Issue 44
  • DOI: 10.1073/pnas.0908848106

Expression of Castor LPAT2 Enhances Ricinoleic Acid Content at the sn-2 Position of Triacylglycerols in Lesquerella Seed
journal, April 2016

  • Chen, Grace; van Erp, Harrie; Martin-Moreno, Jose
  • International Journal of Molecular Sciences, Vol. 17, Issue 4
  • DOI: 10.3390/ijms17040507

Endoplasmic Reticulum-Located PDAT1-2 from Castor Bean Enhances Hydroxy Fatty Acid Accumulation in Transgenic Plants
journal, June 2011

  • Kim, Hyun Uk; Lee, Kyeong-Ryeol; Go, Young Sam
  • Plant and Cell Physiology, Vol. 52, Issue 6
  • DOI: 10.1093/pcp/pcr051

RNA isolation from recalcitrant plant tissue
journal, December 1994

  • Schultz, David J.; Craig, Richard; Cox-Foster, Diana L.
  • Plant Molecular Biology Reporter, Vol. 12, Issue 4
  • DOI: 10.1007/BF02669273

Engineering increased triacylglycerol accumulation in Saccharomyces cerevisiae using a modified type 1 plant diacylglycerol acyltransferase
journal, December 2014

  • Greer, Michael S.; Truksa, Martin; Deng, Wei
  • Applied Microbiology and Biotechnology, Vol. 99, Issue 5
  • DOI: 10.1007/s00253-014-6284-4

Imaging heterogeneity of membrane and storage lipids in transgenic Camelina sativa seeds with altered fatty acid profiles
journal, July 2013

  • Horn, Patrick J.; Silva, Jillian E.; Anderson, Danielle
  • The Plant Journal
  • DOI: 10.1111/tpj.12278

Works referencing / citing this record:

Enhanced Production of High‐Value Cyclopropane Fatty Acid in Yeast Engineered for Increased Lipid Synthesis and Accumulation
journal, October 2018

  • Peng, Huadong; He, Lizhong; Haritos, Victoria S.
  • Biotechnology Journal, Vol. 14, Issue 4
  • DOI: 10.1002/biot.201800487