Novel Lysophospholipid Acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b Responsible for Generation of Palmitate-Docosahexaenoate-Phosphatidylcholine and Phosphatidylethanolamine

Abe, Eriko; Ikeda, Kazutaka; Nutahara, Eri; Hayashi, Masahiro; Yamashita, Atsushi; Taguchi, Ryo; Doi, Kosaku; Honda, Daiske; Okino, Nozomu; Ito, Makoto

doi:10.1371/journal.pone.0102377

Cited by 16 publications

(18 citation statements)

References 38 publications

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“…A. limacinum F26-b, isolated from fallen leaves of Rhizophora mucronata collected at Ishigaki island, Okinawa, Japan, was identified by 18S rRNA gene analysis (11). It was grown in glucose-yeast extract (GY) medium [3% glucose and 1% yeast extract in 1.75% artificial sea water (SEALIFE)] with a 0.1% vitamin mixture (0.2% vitamin B1, 0.001% vitamin B2, and 0.001% vitamin B12) and 0.2% trace elements (3% EDTA di-sodium, 0.15% FeCl 3…”

Section: Strains and Culturementioning

confidence: 99%

“…Genomic DNA was prepared from A. limacinum F26-b using a previously described method (11). The open reading frame encoding 395 amino acids of TLDP1 was cloned from the genomic DNA of A. limacinum F26-b by PCR using the primer set of ADRP-F and ADRP-R (supplemental Table S1).…”

Section: Cloning Of Tldp1 and The Preparation Of Recombinant Tldp1mentioning

confidence: 99%

“…Briefly, a BglII site was added to the tldp1 KO construct by PCRbased site-directed mutagenesis using the primers ADRP-BglII-F and ADRP-BglII-R (supplemental Table S1). The hygromycinresistant gene expression cassette (13) was inserted into the BglII site of the vector according to a previously described method (11). The KO construct was amplified by PCR using the primers ADRP-F and ADRP-R and introduced into A. limacinum F26-b by electroporation with Gene Pulser Xcell.…”

Section: Generation Of Tldp1-disrupted Mutants (Ko)mentioning

confidence: 99%

“…The novel thraustochytrid-specific lipid droplet protein responsible for DHA and DHA-containing phospholipids have been conducted for more than a decade (9)(10)(11)(12)(13)(14)(15)(16) and thraustochytrids have emerged as a useful model for studying the metabolism and accumulation of DHA-containing lipids. We also reported the lipid profile of Aurantiochytrium limacinum F26-b (17), the strain used in this study [the former name of this strain was Schizochytrium sp.…”

mentioning

confidence: 99%

“…We also reported the lipid profile of Aurantiochytrium limacinum F26-b (17), the strain used in this study [the former name of this strain was Schizochytrium sp. F26-b and it was renamed A. limacinum F26-b based on several taxonomic methods, including 18S rRNA gene analysis (11)].…”

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confidence: 99%

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Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b

Watanabe

Sakiyama

Iimi

et al. 2017

Journal of Lipid Research

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Thraustochytrids are marine single-cell protists that produce large amounts of PUFAs, such as DHA. They accumulate PUFAs in lipid droplets (LDs), mainly as constituent(s) of triacylglycerol (TG). We identified a novel protein in the LD fraction of F26-b using 2D-difference gel electrophoresis. The protein clustered with orthologs of thraustochytrids; however, the cluster was evolutionally different from known PAT family proteins or plant LD protein; thus, we named it thraustochytrid-specific LD protein 1 (TLDP1). TLDP1 surrounded LDs when expressed as a GFP-tagged form. Disruption of the gene decreased the content of TG and number of LDs per cell; however, irregular and unusually large LDs were generated in -deficient mutants. Although the level of TG synthesis was unchanged by the disruption of, the level of TG degradation was higher in -deficient mutants than in the WT. These phenotypic abnormalities in-deficient mutants were restored by the expression of These results indicate that TLDP1 is a thraustochytrid-specific LD protein and regulates the TG accumulation and LD morphology in F26-b.

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Section: Strains and Culturementioning

confidence: 99%

Section: Cloning Of Tldp1 and The Preparation Of Recombinant Tldp1mentioning

confidence: 99%

Section: Generation Of Tldp1-disrupted Mutants (Ko)mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b

Watanabe

Sakiyama

Iimi

et al. 2017

Journal of Lipid Research

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Mechanisms of fatty acid synthesis in marine fungus-like protists

Xie

Wang

2015

Appl Microbiol Biotechnol

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Thraustochytrids are unicellular fungus-like protists and are well known for their ability to produce interesting nutraceutical compounds. Significant efforts have been made to improve their efficient production of important fatty acids (FAs), mostly by optimizing fermentation conditions and selecting highly productive thraustochytrid strains. Furthermore, noticeable improvements have been made in understanding the mechanism of FA biosynthesis, allowing for a better understanding of how thraustochytrids assemble these unique metabolites and how their biosynthesis is coupled with other related pathways. This review summarizes recent achievements on two major FA biosynthesis pathways, the standard pathway and the polyketide synthase pathway, and detail features of individual enzymes involved in FA biosynthesis, biotechnological advances in pathway engineering and enzyme characterization, and the discovery of other pathways that affect the efficiency of FA accumulation. Perspectives of biotechnological potential application of thraustochytrids are also discussed.

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Thraustochytrids as production organisms for docosahexaenoic acid (DHA), squalene, and carotenoids

Aasen

Ertesvåg

Heggeset

et al. 2016

Appl Microbiol Biotechnol

157

112

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Thraustochytrids have been applied for industrial production of the omega-3 fatty acid docosahexaenoic (DHA) since the 1990s. During more than 20 years of research on this group of marine, heterotrophic microorganisms, considerable increases in DHA productivities have been obtained by process and medium optimization. Strains of thraustochytrids also produce high levels of squalene and carotenoids, two other commercially interesting compounds with a rapidly growing market potential, but where yet few studies on process optimization have been reported. Thraustochytrids use two pathways for fatty acid synthesis. The saturated fatty acids are produced by the standard fatty acid synthesis, while DHA is synthesized by a polyketide synthase. However, fundamental knowledge about the relationship between the two pathways is still lacking. In the present review, we extract main findings from the high number of reports on process optimization for DHA production and interpret these in the light of the current knowledge of DHA synthesis in thraustochytrids and lipid accumulation in oleaginous microorganisms in general. We also summarize published reports on squalene and carotenoid production and review the current status on strain improvement, which has been hampered by the yet very few published genome sequences and the lack of tools for gene transfer to the organisms. As more sequences now are becoming available, targets for strain improvement can be identified and open for a system-level metabolic engineering for improved productivities.

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Novel Lysophospholipid Acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b Responsible for Generation of Palmitate-Docosahexaenoate-Phosphatidylcholine and Phosphatidylethanolamine

Cited by 16 publications

References 38 publications

Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b

Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b

Mechanisms of fatty acid synthesis in marine fungus-like protists

Thraustochytrids as production organisms for docosahexaenoic acid (DHA), squalene, and carotenoids

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