Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase
Abstract
Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) synthesizes the unusually structured 3-acetyl-1,2-diacylglycerols (acetyl-TAG) found in the seeds of a few plant species. A member of the membrane-bound O-acyltransferase (MBOAT) family, EaDAcT transfers the acetyl group from acetyl-CoA to sn-1,2-diacylglycerol (DAG) to produce acetyl-TAG. In vitro assays demonstrated that the enzyme is also able to utilize butyryl-CoA and hexanoyl-CoA as acyl donors, though with much less efficiency compared with acetyl-CoA. Acyl-CoAs longer than eight carbons were not used by EaDAcT. This extreme substrate specificity of EaDAcT distinguishes it from all other MBOATs which typically catalyze the transfer of much longer acyl groups. In vitro selectivity experiments revealed that EaDAcT preferentially acetylated DAG molecules containing more double bonds over those with less. However, the enzyme was also able to acetylate saturated DAG containing medium chain fatty acids, albeit with less efficiency. Interestingly, EaDAcT could only acetylate the free hydroxyl group of sn-1,2-DAG but not the available hydroxyl groups in sn-1,3-DAG or in monoacylglycerols (MAG). Consistent with its similarity to the jojoba wax synthase, EaDAcT could acetylate fatty alcohols in vitro to produce alkyl acetates. Likewise, when coexpressed in yeast with a fatty acyl-CoA reductase capable of producing fatty alcohols, EaDAcT synthesized alkyl acetates althoughmore »
- Authors:
-
- Kansas State Univ., Manhattan, KS (United States)
- Publication Date:
- Research Org.:
- Colorado State Univ., Fort Collins, CO (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1360143
- Grant/Contract Number:
- SC0012459
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Bioscience Reports
- Additional Journal Information:
- Journal Volume: 36; Journal Issue: 6; Journal ID: ISSN 0144-8463
- Publisher:
- Portland Press - Biochemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; acetyltransferase; MBOAT; triacylglycerols; acetyl‐TAGs; substrate specificity
Citation Formats
Bansal, Sunil, and Durrett, Timothy P. Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase. United States: N. p., 2016.
Web. doi:10.1042/BSR20160277.
Bansal, Sunil, & Durrett, Timothy P. Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase. United States. https://doi.org/10.1042/BSR20160277
Bansal, Sunil, and Durrett, Timothy P. Tue .
"Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase". United States. https://doi.org/10.1042/BSR20160277. https://www.osti.gov/servlets/purl/1360143.
@article{osti_1360143,
title = {Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase},
author = {Bansal, Sunil and Durrett, Timothy P.},
abstractNote = {Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) synthesizes the unusually structured 3-acetyl-1,2-diacylglycerols (acetyl-TAG) found in the seeds of a few plant species. A member of the membrane-bound O-acyltransferase (MBOAT) family, EaDAcT transfers the acetyl group from acetyl-CoA to sn-1,2-diacylglycerol (DAG) to produce acetyl-TAG. In vitro assays demonstrated that the enzyme is also able to utilize butyryl-CoA and hexanoyl-CoA as acyl donors, though with much less efficiency compared with acetyl-CoA. Acyl-CoAs longer than eight carbons were not used by EaDAcT. This extreme substrate specificity of EaDAcT distinguishes it from all other MBOATs which typically catalyze the transfer of much longer acyl groups. In vitro selectivity experiments revealed that EaDAcT preferentially acetylated DAG molecules containing more double bonds over those with less. However, the enzyme was also able to acetylate saturated DAG containing medium chain fatty acids, albeit with less efficiency. Interestingly, EaDAcT could only acetylate the free hydroxyl group of sn-1,2-DAG but not the available hydroxyl groups in sn-1,3-DAG or in monoacylglycerols (MAG). Consistent with its similarity to the jojoba wax synthase, EaDAcT could acetylate fatty alcohols in vitro to produce alkyl acetates. Likewise, when coexpressed in yeast with a fatty acyl-CoA reductase capable of producing fatty alcohols, EaDAcT synthesized alkyl acetates although the efficiency of production was low. As a result, this improved understanding of EaDAcT specificity confirms that the enzyme preferentially utilizes acetyl-CoA to acetylate sn-1,2-DAGs and will be helpful in engineering the production of acetyl-TAG with improved functionality in transgenic plants.},
doi = {10.1042/BSR20160277},
journal = {Bioscience Reports},
number = 6,
volume = 36,
place = {United States},
year = {Tue Nov 08 00:00:00 EST 2016},
month = {Tue Nov 08 00:00:00 EST 2016}
}
Web of Science
Works referenced in this record:
A distinct DGAT with sn-3 acetyltransferase activity that synthesizes unusual, reduced-viscosity oils in Euonymus and transgenic seeds
journal, May 2010
- Durrett, T. P.; McClosky, D. D.; Tumaney, A. W.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 20, p. 9464-9469
Asymmetric triglycerides from Impatiens edgeworthii seed oil
journal, June 1967
- Bagby, M. O.; Smith, C. R.
- Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, Vol. 137, Issue 3
(S)-1,2-diacyl-3-acetins: Optically active triglycerides fromEuonymus verrucosus seed oil
journal, November 1967
- Kleiman, R.; Miller, R. W.; Earle, F. R.
- Lipids, Vol. 2, Issue 6
Positional-Species Composition of Triacylglycerols from the Arils of Mature Euonymus Fruits
journal, October 2014
- Sidorov, Roman A.; Pchelkin, Vasily P.; Zhukov, Anatoly V.
- Journal of the American Oil Chemists' Society, Vol. 91, Issue 12
Field production, purification and analysis of high-oleic acetyl-triacylglycerols from transgenic Camelina sativa
journal, March 2015
- Liu, Jinjie; Tjellström, Henrik; McGlew, Kathleen
- Industrial Crops and Products, Vol. 65
Membrane-bound O-acyltransferases (MBOATs)
journal, June 2011
- Chang, Catherine C. Y.; Sun, Jie; Chang, Ta-Yuan
- Frontiers in Biology, Vol. 6, Issue 3
A superfamily of membrane-bound O -acyltransferases with implications for Wnt signaling
journal, March 2000
- Hofmann, Kay
- Trends in Biochemical Sciences, Vol. 25, Issue 3
Diacylglycerol acyltransferase: A key mediator of plant triacylglycerol synthesis
journal, December 2006
- Lung, Shiu-Cheung; Weselake, Randall J.
- Lipids, Vol. 41, Issue 12
Monounsaturated Fatty Acid Modification of Wnt Protein: Its Role in Wnt Secretion
journal, December 2006
- Takada, Ritsuko; Satomi, Yoshinori; Kurata, Tomoko
- Developmental Cell, Vol. 11, Issue 6
Metabolic engineering of oilseed crops to produce high levels of novel acetyl glyceride oils with reduced viscosity, freezing point and calorific value
journal, March 2015
- Liu, Jinjie; Rice, Adam; McGlew, Kathleen
- Plant Biotechnology Journal, Vol. 13, Issue 6
Large-scale mutational analysis of Kv11.1 reveals molecular insights into type 2 long QT syndrome
journal, November 2014
- Anderson, Corey L.; Kuzmicki, Catherine E.; Childs, Ryan R.
- Nature Communications, Vol. 5, Issue 1
The Yeast ATF1 Acetyltransferase Efficiently Acetylates Insect Pheromone Alcohols: Implications for the Biological Production of Moth Pheromones
journal, January 2016
- Ding, Bao-Jian; Lager, Ida; Bansal, Sunil
- Lipids, Vol. 51, Issue 4
Rapid Quantification of Low-Viscosity Acetyl-Triacylglycerols Using Electrospray Ionization Mass Spectrometry
journal, August 2016
- Bansal, Sunil; Durrett, Timothy P.
- Lipids, Vol. 51, Issue 9
Storage Lipid Synthesis Is Non-essential in Yeast
journal, December 2001
- Sandager, Line; Gustavsson, Maria H.; Ståhl, Ulf
- Journal of Biological Chemistry, Vol. 277, Issue 8, p. 6478-6482
Analysis of Acyl Fluxes through Multiple Pathways of Triacylglycerol Synthesis in Developing Soybean Embryos
journal, March 2009
- Bates, Philip D.; Durrett, Timothy P.; Ohlrogge, John B.
- Plant Physiology, Vol. 150, Issue 1
A fatty acyl-CoA reductase highly expressed in the head of honey bee (Apis mellifera) involves biosynthesis of a wide range of aliphatic fatty alcohols
journal, September 2010
- Teerawanichpan, Prapapan; Robertson, Albert J.; Qiu, Xiao
- Insect Biochemistry and Molecular Biology, Vol. 40, Issue 9
Oil content of Arabidopsis seeds: The influence of seed anatomy, light and plant-to-plant variation
journal, May 2006
- Li, Yonghua; Beisson, Fred; Pollard, Mike
- Phytochemistry, Vol. 67, Issue 9
Ghrelin octanoylation mediated by an orphan lipid transferase
journal, April 2008
- Gutierrez, J. A.; Solenberg, P. J.; Perkins, D. R.
- Proceedings of the National Academy of Sciences, Vol. 105, Issue 17
Identification of the Acyltransferase that Octanoylates Ghrelin, an Appetite-Stimulating Peptide Hormone
journal, February 2008
- Yang, Jing; Brown, Michael S.; Liang, Guosheng
- Cell, Vol. 132, Issue 3
Positional-Species Composition of Diacylglycerol Acetates from Mature Euonymus Seeds
journal, June 2016
- Sidorov, Roman A.; Pchelkin, Vasily P.; Zhukov, Anatoly V.
- Chemistry & Biodiversity, Vol. 13, Issue 6
The triacylglycerol synthesis enzyme DGAT1 also catalyzes the synthesis of diacylglycerols, waxes, and retinyl esters
journal, April 2005
- Yen, Chi-Liang Eric; Monetti, Mara; Burri, Betty J.
- Journal of Lipid Research, Vol. 46, Issue 7
Neopentyl Polyol Ester Lubricants-Bulk Property Optimization
journal, March 1976
- Niedzielski, Edmund L.
- Industrial & Engineering Chemistry Product Research and Development, Vol. 15, Issue 1
Works referencing / citing this record:
A New Class of Acetyl-TAG Present in Seed Oils of Polygala Species
journal, July 2018
- Durrett, Timothy P.
- European Journal of Lipid Science and Technology, Vol. 120, Issue 9
Overexpression of diacylglycerol acetyltransferase from Euonymus europaeus in Yarrowia lipolytica leads to the production of single‐cell oil enriched with 3‐acetyl‐1,2‐diacylglycerols
journal, November 2019
- Gajdoš, Peter; Hambalko, Jaroslav; Nicaud, Jean‐Marc
- Yeast, Vol. 37, Issue 1
Towards the synthetic design of camelina oil enriched in tailored acetyl-triacylglycerols with medium-chain fatty acids
journal, July 2018
- Bansal, Sunil; Kim, Hae Jin; Na, GunNam
- Journal of Experimental Botany, Vol. 69, Issue 18
Membrane topology and identification of key residues of Ea DAcT, a plant MBOAT with unusual substrate specificity
journal, August 2017
- Tran, Tam N. T.; Shelton, Jennifer; Brown, Susan
- The Plant Journal, Vol. 92, Issue 1