Production of wax esters in plant seed oils by oleosomal cotargeting of biosynthetic enzymes

Heilmann, Mareike; Iven, Tim; Ahmann, Katharina; Hornung, Ellen; Stymne, Sten; Feußner, Ivo

doi:10.1194/jlr.m029512

Cited by 48 publications

(93 citation statements)

References 21 publications

(40 reference statements)

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“…In addition to the examples that we detail, the production of many other unusual lipids, such as wax esters and epoxy-fatty acids, has been demonstrated in Arabidopsis [84,85]. However, the development of an effective transformation method has allowed transgenic Camelina lines to be developed almost as easily as with Arabidopsis.…”

Section: Resultsmentioning

confidence: 93%

Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids

Bansal

Durrett

2016

Biochimie

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Triacylglycerols (TAG) containing modified fatty acids with functionality beyond those found in commercially grown oil seed crops can be used as feedstocks for biofuels and bio-based materials. Over the years, advances have been made in transgenically engineering the production of various modified fatty acids in the model plant Arabidopsis thaliana. However, the inability to produce large quantities of transgenic seed has limited the functional testing of the modified oil. In contrast, the emerging oil seed crop Camelina sativa possesses important agronomic traits that recommend it as an ideal production platform for biofuels and industrial feedstocks. Camelina possesses low water and fertilizer requirements and is capable of yields comparable to other oil seed crops, particularly under stress conditions. Importantly, its relatively short growing season enables it to be grown as part of a double cropping system. In addition to these valuable agronomic features, Camelina is amenable to rapid metabolic engineering. The development of a simple and effective transformation method, combined with the availability of abundant transcriptomic and genomic data, has allowed the generation of transgenic Camelina lines capable of synthesizing high levels of unusual lipids. In some cases these levels have surpassed what was achieved in Arabidopsis. Further, the ability to use Camelina as a crop production system has allowed for the large scale growth of transgenic oil seed crops, enabling subsequent physical property testing. The application of new techniques such as genome editing will further increase the suitability of Camelina as an ideal platform for the production of biofuels and bio-materials.

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Section: Resultsmentioning

confidence: 93%

Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids

Bansal

Durrett

2016

Biochimie

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“…Primers used are listed in Supplemental Table S3. DGAT1, PDAT, and amiRNA sequences were cloned into modified pUC18ENTR2 vectors (Heilmann et al, 2012) with napin as a promotor. Final constructs were made by combining four different pUC18ENTR2 vectors (empty or containing a gene/amiRNA) using Gateway technology (Invitrogen) with a modified pCAMBIA33.1 vector (Hornung et al, 2005).…”

Section: Plasmid Constructionmentioning

confidence: 99%

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

et al. 2017

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ORCID IDs: 0000-0001-6929-7859 (S.M.); 0000-0003-3152-0394 (D.S.); 0000-0001-8255-3255 (C.H.); 0000-0003-0489-3072 (K.C.); 0000-0002-9888-7003 (I.F.).Acyltransferases are key contributors to triacylglycerol (TAG) synthesis and, thus, are of great importance for seed oil quality. The effects of increased or decreased expression of ACYL-COENZYME A:DIACYLGLYCEROL ACYLTRANSFERASE1 (DGAT1) or PHOSPHOLIPID:DIACYLGLYCEROL ACYLTRANSFERASE (PDAT) on seed lipid composition were assessed in several Camelina sativa lines. Furthermore, in vitro assays of acyltransferases in microsomal fractions prepared from developing seeds of some of these lines were performed. Decreased expression of DGAT1 led to an increased percentage of 18:3n-3 without any change in total lipid content of the seed. The tri-18:3 TAG increase occurred predominantly in the cotyledon, as determined with matrix-assisted laser desorption/ionization-mass spectrometry, whereas species with two 18:3n-3 acyl groups were elevated in both cotyledon and embryonal axis. PDAT overexpression led to a relative increase of 18:2n-6 at the expense of 18:3n-3, also without affecting the total lipid content. Differential distributions of TAG species also were observed in different parts of the seed. The microsomal assays revealed that C. sativa seeds have very high activity of diacylglycerol-phosphatidylcholine interconversion. The combination of analytical and biochemical data suggests that the higher 18:2n-6 content in the seed oil of the PDAT overexpressors is due to the channeling of fatty acids from phosphatidylcholine into TAG before being desaturated to 18:3n-3, caused by the high activity of PDAT in general and by PDAT specificity for 18:2n-6. The higher levels of 18:3n-3 in DGAT1-silencing lines are likely due to the compensatory activity of a TAG-synthesizing enzyme with specificity for this acyl group and more desaturation of acyl groups occurring on phosphatidylcholine.

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“…Coexpression of the jojoba FAR and WS genes with a jojoba fatty acid elongase (ScFAE) in Arabidopsis was reported to result in significant accumulation of wax esters in seed oil (Lardizabal et al, 2000), however, this promising result was not followed on by the authors. More recently, Heilmann et al co-expressed an endoplasmic reticulum (ER)-localized mouse WS with a mouse peroxisomal FAR re-targeted to the ER to generate wax esters in Arabidopsis seeds with principally C18 and C20 fatty acid and fatty alcohol components (Heilmann et al, 2012). The authors reported that linking theses enzymes at their amino-termini to oleosin, an oil body structural protein, wax ester contents as high as 45 μg/mg seed weight or ∼15% of the total seed oil were achieved.…”

Section: Fish Oil (Omega-3 Lc-pufas) Production In Camelinamentioning

confidence: 99%

Oleaginous crops as integrated production platforms for food, feed, fuel and renewable industrial feedstock

Beaudoin

Sayanova

Haslam

et al. 2014

OCL

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-The world faces considerable challenges including how to produce more biomass for food, feed, fuel and industrial feedstock without significantly impacting on our environment or increasing our consumption of limited resources such as water or petroleum-derived carbon. This has been described as sustainable intensification. Oleaginous crops have the potential to provide renewable resources for all these commodities, provided they can be engineered to meet end-use requirements, and that they can be produced on sufficient scale to meet current growing world population and industrial demand. Although traditional breeding methods have been used successfully to modify the fatty acid composition of oils, metabolic engineering provides a more rapid and direct method for manipulating plant lipid composition. Recent advances in our understanding of the biochemical mechanisms of seed oil biogenesis and the cloning of genes involved in fatty acid and oil metabolic pathways, have allowed the generation of oilseed crops that produce 'designer oils' tailored for specific applications and the conversion of high biomass crops into novel oleaginous crops. However, improvement of complex quantitative traits in oilseed crops remains more challenging as the underlying genetic determinants are still poorly understood. Technological advances in sequencing and computing have allowed the development of an association genetics method applicable to crops with complex genomes. Associative transcriptomics approaches and high throughput lipidomic profiling can be used to identify the genetic components controlling quantitative variation for lipid related traits in polyploid crops like oilseed rape and provide molecular tools for marker assisted breeding. In this review we are citing examples of traits with potential for bio-refining that can be harvested as co-products in seeds, but also in non-harvested biomass.Keywords: Crops improvement / oleaginous crops / metabolic engineering / designer oil / molecular breeding / Genome Wide Association Studies / associative transcriptomics / co-products Résumé -Les oléagineux en tant que plateformes intégrées de production pour l'alimentation humaine et animale, les carburants et les matières premières industrielles renouvelables : Manipulation de la composition lipidique de la plante par ingénierie métabolique et nouvelles opportunités d'associations génétiques pour l'amé-lioration des cultures et la valorisation des coproduits. Le monde fait face à des défis considérables, incluant le moyen de produire davantage de biomasse pour l'alimentation humaine et animale, les carburants et les matières premières industrielles, et ce sans impact environnemental significatif ou sans accroître notre consommation de ressources limitées comme l'eau ou le carbone dérivé du pétrole. Cela a été décrit comme de l'intensification durable. Les oléa-gineux disposent du potentiel nécessaire pour fournir des ressources renouvelables pour toutes ces matières premières, sous réserve qu'ils puissent être manipulés ...

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Production of wax esters in plant seed oils by oleosomal cotargeting of biosynthetic enzymes

Cited by 48 publications

References 21 publications

Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids

Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

Oleaginous crops as integrated production platforms for food, feed, fuel and renewable industrial feedstock

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