Cloning and characterization of a novel diacylglycerol acyltransferase from the diatom Phaeodactylum tricornutum

Cui, Yulin; Zheng, Guoting; Lin, Hanzhi; Jiang, Peng; Qin, Song

doi:10.1007/s10811-013-9991-9

Cited by 25 publications

(22 citation statements)

References 10 publications

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“…Unlike the puzzling roles of soluble DGAT in plants, several pieces of strong evidence support the involvement of DGAT3 in TAG biosynthesis in microalgae and diatoms. For instance, the involvement of DGAT3 in TAG biosynthesis in the diatom Phaeodactylum tricornutum was confirmed by heterologous expression in S. cerevisiae mutant H1246 (Cui et al, ). Similarly, a novel DGAT exclusive to green microalgae with moderate similarity to plant DGAT3 was found to participate in the chloroplastidial de novo synthesis of TAG (Bagnato et al, ).…”

Section: Introductionmentioning

confidence: 96%

Properties and Biotechnological Applications of Acyl‐CoA:diacylglycerol Acyltransferase and Phospholipid:diacylglycerol Acyltransferase from Terrestrial Plants and Microalgae

Caldo

Pal‐Nath

et al. 2018

Lipids

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Triacylglycerol (TAG) is the major storage lipid in most terrestrial plants and microalgae, and has great nutritional and industrial value. Since the demand for vegetable oil is consistently increasing, numerous studies have been focused on improving the TAG content and modifying the fatty‐acid compositions of plant seed oils. In addition, there is a strong research interest in establishing plant vegetative tissues and microalgae as platforms for lipid production. In higher plants and microalgae, TAG biosynthesis occurs via acyl‐CoA‐dependent or acyl‐CoA‐independent pathways. Diacylglycerol acyltransferase (DGAT) catalyzes the last and committed step in the acyl‐CoA‐dependent biosynthesis of TAG, which appears to represent a bottleneck in oil accumulation in some oilseed species. Membrane‐bound and soluble forms of DGAT have been identified with very different amino‐acid sequences and biochemical properties. Alternatively, TAG can be formed through acyl‐CoA‐independent pathways via the catalytic action of membrane‐bound phospholipid:diacylglycerol acyltransferase (PDAT). As the enzymes catalyzing the terminal steps of TAG formation, DGAT and PDAT play crucial roles in determining the flux of carbon into seed TAG and thus have been considered as the key targets for engineering oil production. Here, we summarize the most recent knowledge on DGAT and PDAT in higher plants and microalgae, with the emphasis on their physiological roles, structural features, and regulation. The development of various metabolic engineering strategies to enhance the TAG content and alter the fatty‐acid composition of TAG is also discussed.

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Section: Introductionmentioning

confidence: 96%

Properties and Biotechnological Applications of Acyl‐CoA:diacylglycerol Acyltransferase and Phospholipid:diacylglycerol Acyltransferase from Terrestrial Plants and Microalgae

Caldo

Pal‐Nath

et al. 2018

Lipids

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“…Not all the genes tested restored the TAG phenotype, but of those that did, many showed differing substrate specificity for mono-and polyunsaturated acyl-CoA substrates, e.g. O. tauri DGAT2B showed broad substrate specificity, accepting saturated as well as mono-and polyunsaturated acyl-CoAs as substrates [19], while DGAT3 isoform from P. tricornutum, (PtDGAT3) displayed a preference for the incorporation of unsaturated C18-fatty acids [20].…”

Section: Introductionmentioning

confidence: 99%

Overexpression of an endogenous type 2 diacylglycerol acyltransferase in the marine diatom Phaeodactylum tricornutum enhances lipid production and omega-3 Long Chain Polyunsaturated Fatty Acid content

Haslam

Hamilton

Economou

et al. 2020

Preprint

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Background: Oleaginous microalgae represent a valuable resource for the production of high value molecules. Considering the importance of omega-3 long chain polyunsaturated fatty acids (LC-PUFAs) for human health and nutrition the yields of high value eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) require significant improvement to meet demand, however, the current cost of production remains high. A promising approach is to metabolically engineer strains with enhanced levels of triacylglycerols (TAGs) enriched in EPA and DHA.Results: Recently we have engineered the marine diatom Phaeodactylum tricornutum to accumulate enhanced levels of DHA in TAG. To further improve the incorporation of omega-3 LC-PUFAs in TAG we focused our effort on the identification of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT) capable of improving lipid production and the incorporation of DHA in TAG. DGAT is a key enzyme in lipid synthesis. Following a diatom based in vivo screen of candidate DGATs, a native P. tricornutum DGAT2B was taken forward for detailed characterisation. Overexpression of the endogenous P. tricornutum DGAT2B was confirmed by qRT-PCR and the transgenic strain grew successfully in comparison to wildtype. PtDGAT2B has broad substrate specificity with preferences for C16 and LC-PUFAs acyl groups. Moreover, the overexpression of an endogenous DGAT2B resulted in higher lipid yields and enhanced levels of DHA in TAG. Furthermore, a combined overexpression of the endogenous DGAT2B and ectopic expression of a Δ5-elongase showed how iterative metabolic engineering can be used to increase DHA and TAG content, irrespective of nitrogen treatment.Conclusion: This study provides further insight into lipid metabolism in P. tricornutum and suggests a metabolic engineering approach for the efficient production of EPA and DHA in microalgae.

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“…In our previous report, we successfully developed transgenic P. tricornutum with increased lipid accumulation by overexpressing type 2 diacylglycerol acyl transferase (DGAT) [ 19 ]. Similarly, overexpression of DGAT in P. tricornutum resulted in increased proportion of polyunsaturated fatty acids [ 20 , 21 ]. However, no GPAT has been characterized in microalgae.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of a glycerol-3-phosphate acyltransferase reveals key features essential for triacylglycerol production in Phaeodactylum tricornutum

Niu

Wang

et al. 2016

Biotechnol Biofuels

110

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BackgroundThe marine diatom, Phaeodactylum tricornutum, has become a model for studying lipid metabolism and its triacylglycerol (TAG) synthesis pathway makes it an ideal target for metabolic engineering to improve lipid productivity. However, the genetic background and metabolic networks of fatty acid biosynthesis in diatoms are not well understood. Glycerol-3-phosphate acyltransferase (GPAT) is the critical enzyme that catalyzes the first step of TAG formation. So far, characterization of GPAT in marine microalgae has not been reported, especially at the level of comprehensive sequence-structure and functional analysis.ResultsA GPAT was cloned from P. tricornutum and overexpressed in P. tricornutum. Volumes of oil bodies were produced and the neutral lipid content was increased by twofold determined by Nile red fluorescence staining. Fatty acid composition was analyzed by GC–MS, which showed significantly higher proportion of unsaturated fatty acids compared to wild type.ConclusionThese results suggested that the identified GPAT could upregulate TAG biosynthesis in P. tricornutum. Moreover, this study offers insight into the lipid metabolism of diatoms and supports the role of microalgal strains for biofuels production.

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Cloning and characterization of a novel diacylglycerol acyltransferase from the diatom Phaeodactylum tricornutum

Cited by 25 publications

References 10 publications

Properties and Biotechnological Applications of Acyl‐CoA:diacylglycerol Acyltransferase and Phospholipid:diacylglycerol Acyltransferase from Terrestrial Plants and Microalgae

Properties and Biotechnological Applications of Acyl‐CoA:diacylglycerol Acyltransferase and Phospholipid:diacylglycerol Acyltransferase from Terrestrial Plants and Microalgae

Overexpression of an endogenous type 2 diacylglycerol acyltransferase in the marine diatom Phaeodactylum tricornutum enhances lipid production and omega-3 Long Chain Polyunsaturated Fatty Acid content

Molecular characterization of a glycerol-3-phosphate acyltransferase reveals key features essential for triacylglycerol production in Phaeodactylum tricornutum

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