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Mar. Drugs 2014, 12(3), 1317-1334; doi:10.3390/md12031317
Article

Isolation and Characterization of the Diatom Phaeodactylum Δ5-Elongase Gene for Transgenic LC-PUFA Production in Pichia pastoris

1,2,†
, 1,†
, 1,2
, 1,2
, 1
 and 1,2,*
Received: 12 November 2013; in revised form: 13 February 2014 / Accepted: 17 February 2014 / Published: 7 March 2014
(This article belongs to the Special Issue Metabolites in Diatoms)
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Abstract: The diatom Phaeodactylum tricornutum can accumulate eicosapentaenoic acid (EPA) up to 30% of the total fatty acids. This species has been targeted for isolating gene encoding desaturases and elongases for long-chain polyunsaturated fatty acid (LC-PUFA) metabolic engineering. Here we first report the cloning and characterization of Δ5-elongase gene in P. tricornutum. A full-length cDNA sequence, designated PhtELO5, was shown to contain a 1110 bp open reading frame encoding a 369 amino acid polypeptide. The putative protein contains seven transmembrane regions and two elongase characteristic motifs of FLHXYHH and MYSYY, the latter being typical for microalgal Δ5-elongases. Phylogenetic analysis indicated that PhtELO5 belongs to the ELO5 group, tightly clustered with the counterpart of Thalassiosira pseudonana. Heterologous expression of PhtELO5 in Pichia pastoris confirmed that it encodes a specific Δ5-elongase capable of elongating arachidonic acid and eicosapentaenoic acid. Co-expression of PhtELO5 and IsFAD4 (a ∆4-desaturase from Isochrysis sphaerica) demonstrated that the high-efficiency biosynthetic pathway of docosahexaenoic acid was assembled in the transgenic yeast. Substrate competition revealed that PhtELO5 exhibited higher activity towards n-3 PUFA than n-6 PUFA. It is hypothesized that Phaeodactylum ELO5 may preferentially participate in biosynthesis of transgenic LC-PUFA via a n-3 pathway in the yeast host.
Keywords: diatom fatty acids metabolites; Phaeodactylum Δ5-elongase; functional stacking of ELO5 and FAD4 diatom fatty acids metabolites; Phaeodactylum Δ5-elongase; functional stacking of ELO5 and FAD4
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Jiang, M.; Guo, B.; Wan, X.; Gong, Y.; Zhang, Y.; Hu, C. Isolation and Characterization of the Diatom Phaeodactylum Δ5-Elongase Gene for Transgenic LC-PUFA Production in Pichia pastoris. Mar. Drugs 2014, 12, 1317-1334.

AMA Style

Jiang M, Guo B, Wan X, Gong Y, Zhang Y, Hu C. Isolation and Characterization of the Diatom Phaeodactylum Δ5-Elongase Gene for Transgenic LC-PUFA Production in Pichia pastoris. Marine Drugs. 2014; 12(3):1317-1334.

Chicago/Turabian Style

Jiang, Mulan; Guo, Bing; Wan, Xia; Gong, Yangmin; Zhang, Yinbo; Hu, Chuanjiong. 2014. "Isolation and Characterization of the Diatom Phaeodactylum Δ5-Elongase Gene for Transgenic LC-PUFA Production in Pichia pastoris." Mar. Drugs 12, no. 3: 1317-1334.


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