Increase of eicosapentaenoic acid in thraustochytrids through thraustochytrid ubiquitin promoter-driven expression of a fatty acid Δ5 desaturase gene

Takumi Kobayashi, Keishi Sakaguchi, Takanori Matsuda, Eriko Abe, Yoichiro Hama, Masahiro Hayashi, Daiske Honda, Yuji Okita, Shinichi Sugimoto, Nozomu Okino, Makoto Ito

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Thraustochytrids, marine protists known to accumulate polyunsaturated fatty acids (PUFAs) in lipid droplets, are considered an alternative to fish oils as a source of PUFAs. The major fatty acids produced in thraustochytrids are palmitic acid (C16:0), n - 6 docosapentaenoic acid (DPA) (C22:5n - 6 ), and docosahexaenoic acid (DHA) (C22:6n - 3), with eicosapentaenoic acid (EPA) (C20:5n - 3) and arachidonic acid (AA) (C20:4n - 6) as minor constituents. We attempted here to alter the fatty acid composition of thraustochytrids through the expression of a fatty acid Δ5 desaturase gene driven by the thraustochytrid ubiquitin promoter. The gene was functionally expressed in Aurantiochytrium limacinum mh0186, increasing the amount of EPA converted from eicosatetraenoic acid (ETA) (C20:4n - 3) by the Δ5 desaturase. The levels of EPA and AA were also increased by 4.6and 13.2-fold in the transgenic thraustochytrids compared to levels in the mock transfectants when ETA and dihomo-γ-linolenic acid (DGLA) (C20:3n - 6) were added to the culture at 0.1 mM. Interestingly, the amount of EPA in the transgenic thraustochytrids increased in proportion to the amount of ETA added to the culture up to 0.4 mM. The rates of conversion and accumulation of EPA were much higher in the thraustochytrids than in baker's yeasts when the desaturase gene was expressed with the respective promoters. This report describes for the first time the finding that an increase of EPA could be accomplished by introducing the Δ5 desaturase gene into thraustochytrids and indicates that molecular breeding of thraustochytrids is a promising strategy for generating beneficial PUFAs.

Original languageEnglish
Pages (from-to)3870-3876
Number of pages7
JournalApplied and environmental microbiology
Volume77
Issue number11
DOIs
Publication statusPublished - Jun 1 2011

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Fatty Acid Desaturases
Eicosapentaenoic Acid
ubiquitin
eicosapentaenoic acid
Ubiquitin
fatty acid
promoter regions
Arachidonic Acids
fatty acids
gene
acid
Unsaturated Fatty Acids
Genes
polyunsaturated fatty acids
genes
arachidonic acid
Arachidonic Acid
acids
Fatty Acids
DNA Shuffling

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Increase of eicosapentaenoic acid in thraustochytrids through thraustochytrid ubiquitin promoter-driven expression of a fatty acid Δ5 desaturase gene. / Kobayashi, Takumi; Sakaguchi, Keishi; Matsuda, Takanori; Abe, Eriko; Hama, Yoichiro; Hayashi, Masahiro; Honda, Daiske; Okita, Yuji; Sugimoto, Shinichi; Okino, Nozomu; Ito, Makoto.

In: Applied and environmental microbiology, Vol. 77, No. 11, 01.06.2011, p. 3870-3876.

Research output: Contribution to journalArticle

Kobayashi, Takumi ; Sakaguchi, Keishi ; Matsuda, Takanori ; Abe, Eriko ; Hama, Yoichiro ; Hayashi, Masahiro ; Honda, Daiske ; Okita, Yuji ; Sugimoto, Shinichi ; Okino, Nozomu ; Ito, Makoto. / Increase of eicosapentaenoic acid in thraustochytrids through thraustochytrid ubiquitin promoter-driven expression of a fatty acid Δ5 desaturase gene. In: Applied and environmental microbiology. 2011 ; Vol. 77, No. 11. pp. 3870-3876.
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abstract = "Thraustochytrids, marine protists known to accumulate polyunsaturated fatty acids (PUFAs) in lipid droplets, are considered an alternative to fish oils as a source of PUFAs. The major fatty acids produced in thraustochytrids are palmitic acid (C16:0), n - 6 docosapentaenoic acid (DPA) (C22:5n - 6 ), and docosahexaenoic acid (DHA) (C22:6n - 3), with eicosapentaenoic acid (EPA) (C20:5n - 3) and arachidonic acid (AA) (C20:4n - 6) as minor constituents. We attempted here to alter the fatty acid composition of thraustochytrids through the expression of a fatty acid Δ5 desaturase gene driven by the thraustochytrid ubiquitin promoter. The gene was functionally expressed in Aurantiochytrium limacinum mh0186, increasing the amount of EPA converted from eicosatetraenoic acid (ETA) (C20:4n - 3) by the Δ5 desaturase. The levels of EPA and AA were also increased by 4.6and 13.2-fold in the transgenic thraustochytrids compared to levels in the mock transfectants when ETA and dihomo-γ-linolenic acid (DGLA) (C20:3n - 6) were added to the culture at 0.1 mM. Interestingly, the amount of EPA in the transgenic thraustochytrids increased in proportion to the amount of ETA added to the culture up to 0.4 mM. The rates of conversion and accumulation of EPA were much higher in the thraustochytrids than in baker's yeasts when the desaturase gene was expressed with the respective promoters. This report describes for the first time the finding that an increase of EPA could be accomplished by introducing the Δ5 desaturase gene into thraustochytrids and indicates that molecular breeding of thraustochytrids is a promising strategy for generating beneficial PUFAs.",
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