Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase

Bansal, Sunil; Durrett, Timothy P.

doi:10.1042/BSR20160277

Defining the extreme substrate specificity of Euonymus alatus diacylglycerol acetyltransferase, an unusual membrane-bound O-acyltransferase

Journal Article · Tue Nov 08 00:00:00 EST 2016 · Bioscience Reports

DOI:https://doi.org/10.1042/BSR20160277· OSTI ID:1360143

Bansal, Sunil ^[1]; Durrett, Timothy P. ^[2]

Kansas State Univ., Manhattan, KS (United States); Kansas State university
Kansas State Univ., Manhattan, KS (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Colorado State Univ., Fort Collins, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: SC0012459

OSTI ID:: 1360143

Journal Information:: Bioscience Reports, Journal Name: Bioscience Reports Journal Issue: 6 Vol. 36; ISSN 0144-8463

Publisher:: Portland Press - Biochemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

References (22)

Positional-Species Composition of Diacylglycerol Acetates from Mature Euonymus Seeds Sidorov, Roman A.; Pchelkin, Vasily P.; Zhukov, Anatoly V. Chemistry & Biodiversity, Vol. 13, Issue 6 https://doi.org/10.1002/cbdv.201500269	journal	June 2016
(S)-1,2-diacyl-3-acetins: Optically active triglycerides fromEuonymus verrucosus seed oil Kleiman, R.; Miller, R. W.; Earle, F. R. Lipids, Vol. 2, Issue 6 https://doi.org/10.1007/BF02533174	journal	November 1967
Membrane-bound O-acyltransferases (MBOATs) Chang, Catherine C. Y.; Sun, Jie; Chang, Ta-Yuan Frontiers in Biology, Vol. 6, Issue 3 https://doi.org/10.1007/s11515-011-1149-z	journal	June 2011
Diacylglycerol acyltransferase: A key mediator of plant triacylglycerol synthesis Lung, Shiu-Cheung; Weselake, Randall J. Lipids, Vol. 41, Issue 12 https://doi.org/10.1007/s11745-006-5057-y	journal	December 2006
The Yeast ATF1 Acetyltransferase Efficiently Acetylates Insect Pheromone Alcohols: Implications for the Biological Production of Moth Pheromones Ding, Bao-Jian; Lager, Ida; Bansal, Sunil Lipids, Vol. 51, Issue 4 https://doi.org/10.1007/s11745-016-4122-4	journal	January 2016
Rapid Quantification of Low-Viscosity Acetyl-Triacylglycerols Using Electrospray Ionization Mass Spectrometry Bansal, Sunil; Durrett, Timothy P. Lipids, Vol. 51, Issue 9 https://doi.org/10.1007/s11745-016-4179-0	journal	August 2016
Positional-Species Composition of Triacylglycerols from the Arils of Mature Euonymus Fruits Sidorov, Roman A.; Pchelkin, Vasily P.; Zhukov, Anatoly V. Journal of the American Oil Chemists' Society, Vol. 91, Issue 12 https://doi.org/10.1007/s11746-014-2553-8	journal	October 2014
Asymmetric triglycerides from Impatiens edgeworthii seed oil Bagby, M. O.; Smith, C. R. Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, Vol. 137, Issue 3 https://doi.org/10.1016/0005-2760(67)90128-2	journal	June 1967
A superfamily of membrane-bound O -acyltransferases with implications for Wnt signaling Hofmann, Kay Trends in Biochemical Sciences, Vol. 25, Issue 3 https://doi.org/10.1016/S0968-0004(99)01539-X	journal	March 2000
Identification of the Acyltransferase that Octanoylates Ghrelin, an Appetite-Stimulating Peptide Hormone Yang, Jing; Brown, Michael S.; Liang, Guosheng Cell, Vol. 132, Issue 3 https://doi.org/10.1016/j.cell.2008.01.017	journal	February 2008
Monounsaturated Fatty Acid Modification of Wnt Protein: Its Role in Wnt Secretion Takada, Ritsuko; Satomi, Yoshinori; Kurata, Tomoko Developmental Cell, Vol. 11, Issue 6 https://doi.org/10.1016/j.devcel.2006.10.003	journal	December 2006
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 Teerawanichpan, Prapapan; Robertson, Albert J.; Qiu, Xiao Insect Biochemistry and Molecular Biology, Vol. 40, Issue 9 https://doi.org/10.1016/j.ibmb.2010.06.004	journal	September 2010
Field production, purification and analysis of high-oleic acetyl-triacylglycerols from transgenic Camelina sativa Liu, Jinjie; Tjellström, Henrik; McGlew, Kathleen Industrial Crops and Products, Vol. 65 https://doi.org/10.1016/j.indcrop.2014.11.019	journal	March 2015
Oil content of Arabidopsis seeds: The influence of seed anatomy, light and plant-to-plant variation Li, Yonghua; Beisson, Fred; Pollard, Mike Phytochemistry, Vol. 67, Issue 9 https://doi.org/10.1016/j.phytochem.2006.02.015	journal	May 2006
Neopentyl Polyol Ester Lubricants-Bulk Property Optimization Niedzielski, Edmund L. Industrial & Engineering Chemistry Product Research and Development, Vol. 15, Issue 1 https://doi.org/10.1021/i360057a010	journal	March 1976
Large-scale mutational analysis of Kv11.1 reveals molecular insights into type 2 long QT syndrome Anderson, Corey L.; Kuzmicki, Catherine E.; Childs, Ryan R. Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms6535	journal	November 2014
Ghrelin octanoylation mediated by an orphan lipid transferase Gutierrez, J. A.; Solenberg, P. J.; Perkins, D. R. Proceedings of the National Academy of Sciences, Vol. 105, Issue 17 https://doi.org/10.1073/pnas.0800708105	journal	April 2008
A distinct DGAT with sn-3 acetyltransferase activity that synthesizes unusual, reduced-viscosity oils in Euonymus and transgenic seeds Durrett, T. P.; McClosky, D. D.; Tumaney, A. W. Proceedings of the National Academy of Sciences, Vol. 107, Issue 20, p. 9464-9469 https://doi.org/10.1073/pnas.1001707107	journal	May 2010
Storage Lipid Synthesis Is Non-essential in Yeast Sandager, Line; Gustavsson, Maria H.; Ståhl, Ulf Journal of Biological Chemistry, Vol. 277, Issue 8, p. 6478-6482 https://doi.org/10.1074/jbc.M109109200	journal	December 2001
Analysis of Acyl Fluxes through Multiple Pathways of Triacylglycerol Synthesis in Developing Soybean Embryos Bates, Philip D.; Durrett, Timothy P.; Ohlrogge, John B. Plant Physiology, Vol. 150, Issue 1 https://doi.org/10.1104/pp.109.137737	journal	March 2009
Metabolic engineering of oilseed crops to produce high levels of novel acetyl glyceride oils with reduced viscosity, freezing point and calorific value Liu, Jinjie; Rice, Adam; McGlew, Kathleen Plant Biotechnology Journal, Vol. 13, Issue 6 https://doi.org/10.1111/pbi.12325	journal	March 2015
The triacylglycerol synthesis enzyme DGAT1 also catalyzes the synthesis of diacylglycerols, waxes, and retinyl esters Yen, Chi-Liang Eric; Monetti, Mara; Burri, Betty J. Journal of Lipid Research, Vol. 46, Issue 7 https://doi.org/10.1194/jlr.M500036-JLR200	journal	April 2005

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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 Gajdoš, Peter; Hambalko, Jaroslav; Nicaud, Jean‐Marc Yeast, Vol. 37, Issue 1 https://doi.org/10.1002/yea.3442	journal	November 2019
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