Castor diacylglycerol acyltransferase type 1 (DGAT1) displays greater activity with diricinolein than Arabidopsis DGAT1

McKeon, Thomas A.; He, Xiaohua

doi:10.1016/j.bcab.2015.01.005

Cited by 13 publications

(9 citation statements)

References 17 publications

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“…Contrary to the results obtained with the transcription factor WRI1, the enzyme DGAT1 cloned from Arabidopsis was more effective than the one from castor bean (Figures and a). This result is in line with previous finding on the properties of both enzymes, which showed that RcDGAT1 is specialized in forming TAG using hydroxy‐fatty acids, which are expected to be less available in plants other than castor bean (McKeon and He, ). Thus, using a combinatorial approach with genes from different sources was beneficial for TAG accumulation and storage of sesquiterpenes.…”

Section: Discussionsupporting

confidence: 93%

Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves

Delatte

Scaiola

Molenaar

et al. 2018

Plant Biotechnology Journal

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SummaryPlants store volatile compounds in specialized organs. The properties of these storage organs prevent precarious evaporation and protect neighbouring tissues from cytotoxicity. Metabolic engineering of plants is often carried out in tissues such as leaf mesophyll cells, which are abundant and easily accessible by engineering tools. However, these tissues are not suitable for the storage of volatile and hydrophobic compound such as sesquiterpenes and engineered volatiles are often lost into the headspace. In this study, we show that the seeds of Arabidopsis thaliana, which naturally contain lipid bodies, accumulate sesquiterpenes upon engineered expression. Subsequently, storage of volatile sesquiterpenes was achieved in Nicotiana benthamiana leaf tissue, by introducing oleosin‐coated lipid bodies through metabolic engineering. Hereto, different combinations of genes encoding diacylglycerol acyltransferases (DGATs), transcription factors (WRINKL1) and oleosins (OLE1), from the oil seed‐producing species castor bean (Ricinus communis) and Arabidopsis, were assessed for their suitability to promote lipid body formation. Co‐expression of α‐bisabolol synthase with Arabidopsis DGAT1 and WRINKL1 and OLE1 from castor bean promoted storage of α‐bisabolol in N. benthamiana mesophyll tissue more than 17‐fold. A clear correlation was found between neutral lipids and storage of sesquiterpenes, using synthases for α‐bisabolol, (E)‐β‐caryophyllene and α‐barbatene. The co‐localization of neutral lipids and α‐bisabolol was shown using microscopy. This work demonstrates that lipid bodies can be used as intracellular storage compartment for hydrophobic sesquiterpenes, also in the vegetative parts of plants, creating the possibility to improve yields of metabolic engineering strategies in plants.

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Section: Discussionsupporting

confidence: 93%

Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves

Delatte

Scaiola

Molenaar

et al. 2018

Plant Biotechnology Journal

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“…Furthermore, several of the alterations were also present in DGAT1s of castor bean, tung tree and Vernonia, which may reflect evolutionary adaptations of Physaria DGAT1 in response to HFA-rich acyl CoA and DAG pools ( Figure S9). Furthermore, although DGAT2s of castor bean and Vernonia galamensis (which accumulates epoxy FAs) appear to be the major unusual FA-specific acyltransferase, the DGAT1 isoforms of both species have been shown experimentally to have enhanced specificity for unusual FAs relative to species that do not accumulate unusual FAs (Li et al, 2010b;McKeon and He, 2015). These examples of diverged acyltransferase substrate specificity, expression levels and sequence alterations probably illustrate alternative co-evolution strategies in different species to maximize accumulation and accommodate HFA.…”

Section: Modified Acyltransferases May Have Complementary Roles In Hfmentioning

confidence: 99%

Identification of multiple lipid genes with modifications in expression and sequence associated with the evolution of hydroxy fatty acid accumulation in Physaria fendleri

et al. 2016

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Two Brassicaceae species, Physaria fendleri and Camelina sativa, are genetically very closely related to each other and to Arabidopsis thaliana. Physaria fendleri seeds contain over 50% hydroxy fatty acids (HFAs), while Camelina sativa and Arabidopsis do not accumulate HFAs. To better understand how plants evolved new biochemical pathways with the capacity to accumulate high levels of unusual fatty acids, transcript expression and protein sequences of developing seeds of Physaria fendleri, wild-type Camelina sativa, and Camelina sativa expressing a castor bean (Ricinus communis) hydroxylase were analyzed. A number of potential evolutionary adaptations within lipid metabolism that probably enhance HFA production and accumulation in Physaria fendleri, and, in their absence, limit accumulation in transgenic tissues were revealed. These adaptations occurred in at least 20 genes within several lipid pathways from the onset of fatty acid synthesis and its regulation to the assembly of triacylglycerols. Lipid genes of Physaria fendleri appear to have co-evolved through modulation of transcriptional abundances and alterations within protein sequences. Only a handful of genes showed evidence for sequence adaptation through gene duplication. Collectively, these evolutionary changes probably occurred to minimize deleterious effects of high HFA amounts and/or to enhance accumulation for physiological advantage. These results shed light on the evolution of pathways for novel fatty acid production in seeds, help explain some of the current limitations to accumulation of HFAs in transgenic plants, and may provide improved strategies for future engineering of their production.

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“…Acyl-editing and TAG assembly processes play a pivotal role in the enrichment of unusual fatty acid in higher plants (Figure A). − The lack of such a specialized metabolic network in yeast could cause the retention of PuA in PC, which may trigger potential feedback inhibition and reduce the accumulation of PuA in storage lipids. To test the functions of related genes in PuA synthesis in yeast, five putative acyl-channeling genes from pomegranate, including PLA 2 , LPCAT , DGAT2 , PDCT , and PDAT , were synthesized and cloned into pESC-Leu in pairs.…”

Section: Resultsmentioning

confidence: 99%

“…The efficient synthesis and channeling of PuA and other unusual fatty acids in plants that can accumulate large amounts of these fatty acids often requires the contribution of enzymes with special substrate specificities and selectivities. Some such enzymes have been reported in acyl-editing, lipid biosynthetic, and lipid regulatory steps, including phospholipase A 2 (PLA 2 ), lysophospholipid acyltransferase (LPCAT), phosphatidylcholine: diacylglycerol cholinephosphotransferase (PDCT), phospholipid: diacylglycerol acyltransferase (PDAT), and acyl-CoA: diacylglycerol acyltransferase (DGAT). ,− These acyl-editing enzymes may be crucial for enhancing PuA assembly into TAG and thus provide valuable candidates for engineering PuA production. In addition to manipulating TAG assembly (“Pull”), other metabolic engineering strategies for increasing PuA production in microorganisms include increasing fatty acid biosynthesis (“Push”) and preventing TAG turnover (“Protect”). , …”

Section: Introductionmentioning

confidence: 99%

Improving the Production of Punicic Acid in Baker’s Yeast by Engineering Genes in Acyl Channeling Processes and Adjusting Precursor Supply

Wang

Holic̆

et al. 2021

J. Agric. Food Chem.

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Punicic acid (PuA) is a high-value edible conjugated fatty acid with strong bioactivities and has important potential applications in nutraceutical, pharmaceutical, feeding, and oleochemical industries. Since the production of PuA is severely limited by the fact that its natural source (pomegranate seed oil) is not readily available on a large scale, there is considerable interest in understanding the biosynthesis and accumulation of this plant-based unusual fatty acid in transgenic microorganisms to support the rational design of biotechnological approaches for PuA production via fermentation. Here, we tested the effectiveness of genetic engineering and precursor supply in PuA production in the model yeast strain Saccharomyces cerevisiae. The results revealed that the combination of precursor feeding and co-expression of selected genes in acyl channeling processes created an effective "push−pull" approach to increase PuA content, which could prove valuable in future efforts to produce PuA in industrial yeast and other microorganisms via fermentation.

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Castor diacylglycerol acyltransferase type 1 (DGAT1) displays greater activity with diricinolein than Arabidopsis DGAT1

Cited by 13 publications

References 17 publications

Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves

Engineering storage capacity for volatile sesquiterpenes in Nicotiana benthamiana leaves

Identification of multiple lipid genes with modifications in expression and sequence associated with the evolution of hydroxy fatty acid accumulation in Physaria fendleri

Improving the Production of Punicic Acid in Baker’s Yeast by Engineering Genes in Acyl Channeling Processes and Adjusting Precursor Supply

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