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 language | English |
|---|---|
| Pages (from-to) | 3870-3876 |
| Number of pages | 7 |
| Journal | Applied and environmental microbiology |
| Volume | 77 |
| Issue number | 11 |
| DOIs | |
| Publication status | Published - Jun 1 2011 |
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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 journal › Article
}
TY - JOUR
T1 - Increase of eicosapentaenoic acid in thraustochytrids through thraustochytrid ubiquitin promoter-driven expression of a fatty acid Δ5 desaturase gene
AU - Kobayashi, Takumi
AU - Sakaguchi, Keishi
AU - Matsuda, Takanori
AU - Abe, Eriko
AU - Hama, Yoichiro
AU - Hayashi, Masahiro
AU - Honda, Daiske
AU - Okita, Yuji
AU - Sugimoto, Shinichi
AU - Okino, Nozomu
AU - Ito, Makoto
PY - 2011/6/1
Y1 - 2011/6/1
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=79958284114&partnerID=8YFLogxK
U2 - 10.1128/AEM.02664-10
DO - 10.1128/AEM.02664-10
M3 - Article
C2 - 21478316
AN - SCOPUS:79958284114
VL - 77
SP - 3870
EP - 3876
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
SN - 0099-2240
IS - 11
ER -