Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304
Thraustochytrids are known to synthesize PUFAs such as docosahexaenoic acid (DHA). Accumulating evidence suggests the presence of two synthetic pathways of PUFAs in thraustochytrids: the polyketide synthase-like (PUFA synthase) and desaturase/elongase (standard) pathways. It remains unclear whether...
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2012-06-01
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doaj-c4e83a015c234f9b934a5fe8c2a655cc2021-04-28T05:57:49ZengElsevierJournal of Lipid Research0022-22752012-06-0153612101222Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304Takanori Matsuda0Keishi Sakaguchi1Rie Hamaguchi2Takumi Kobayashi3Eriko Abe4Yoichiro Hama5Masahiro Hayashi6Daiske Honda7Yuji Okita8Shinichi Sugimoto9Nozomu Okino10Makoto Ito11Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanDepartment of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanDepartment of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanDepartment of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanDepartment of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanFaculty of Agriculture, Saga University, 1 Honjo, Saga 840-8502, JapanFaculty of Agriculture, Miyazaki University, Miyazaki 889-2192, JapanFaculty of Science and Engineering, Konan University, Okamoto 8-9-1 Higashinada-ku Kobe 658-8501, JapanNippon Suisan Kaisha, Ltd., 6-2 Otemachi 2-chome, Chiyoda-ku, Tokyo 100-8686, Japan;Nippon Suisan Kaisha, Ltd., 6-2 Otemachi 2-chome, Chiyoda-ku, Tokyo 100-8686, Japan;Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanTo whom correspondence should be addressed.; Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan; Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan; To whom correspondence should be addressed.Thraustochytrids are known to synthesize PUFAs such as docosahexaenoic acid (DHA). Accumulating evidence suggests the presence of two synthetic pathways of PUFAs in thraustochytrids: the polyketide synthase-like (PUFA synthase) and desaturase/elongase (standard) pathways. It remains unclear whether the latter pathway functions in thraustochytrids. In this study, we report that the standard pathway produces PUFA in Thraustochytrium aureum ATCC 34304. We isolated a gene encoding a putative Δ12-fatty acid desaturase (TauΔ12des) from T. aureum. Yeasts transformed with the tauΔ12des converted endogenous oleic acid (OA) into linoleic acid (LA). The disruption of the tauΔ12des in T. aureum by homologous recombination resulted in the accumulation of OA and a decrease in the levels of LA and its downstream PUFAs. However, the DHA content was increased slightly in tauΔ12des-disruption mutants, suggesting that DHA is primarily produced in T. aureum via the PUFA synthase pathway. The transformation of the tauΔ12des-disruption mutants with a tauΔ12des expression cassette restored the wild-type fatty acid profiles. These data clearly indicate that TauΔ12des functions as Δ12-fatty acid desaturase in the standard pathway of T. aureum and demonstrate that this thraustochytrid produces PUFAs via both the PUFA synthase and the standard pathways.http://www.sciencedirect.com/science/article/pii/S0022227520317144polyunsaturated fatty acidpolyketide synthase-like (PUFA synthase) pathwaydesaturase/elongase (standard) pathwayhomologous recombinationtargeted gene disruption |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Takanori Matsuda Keishi Sakaguchi Rie Hamaguchi Takumi Kobayashi Eriko Abe Yoichiro Hama Masahiro Hayashi Daiske Honda Yuji Okita Shinichi Sugimoto Nozomu Okino Makoto Ito |
spellingShingle |
Takanori Matsuda Keishi Sakaguchi Rie Hamaguchi Takumi Kobayashi Eriko Abe Yoichiro Hama Masahiro Hayashi Daiske Honda Yuji Okita Shinichi Sugimoto Nozomu Okino Makoto Ito Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 Journal of Lipid Research polyunsaturated fatty acid polyketide synthase-like (PUFA synthase) pathway desaturase/elongase (standard) pathway homologous recombination targeted gene disruption |
author_facet |
Takanori Matsuda Keishi Sakaguchi Rie Hamaguchi Takumi Kobayashi Eriko Abe Yoichiro Hama Masahiro Hayashi Daiske Honda Yuji Okita Shinichi Sugimoto Nozomu Okino Makoto Ito |
author_sort |
Takanori Matsuda |
title |
Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 |
title_short |
Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 |
title_full |
Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 |
title_fullStr |
Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 |
title_full_unstemmed |
Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304 |
title_sort |
analysis of δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of pufas in t. aureum atcc 34304 |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
2012-06-01 |
description |
Thraustochytrids are known to synthesize PUFAs such as docosahexaenoic acid (DHA). Accumulating evidence suggests the presence of two synthetic pathways of PUFAs in thraustochytrids: the polyketide synthase-like (PUFA synthase) and desaturase/elongase (standard) pathways. It remains unclear whether the latter pathway functions in thraustochytrids. In this study, we report that the standard pathway produces PUFA in Thraustochytrium aureum ATCC 34304. We isolated a gene encoding a putative Δ12-fatty acid desaturase (TauΔ12des) from T. aureum. Yeasts transformed with the tauΔ12des converted endogenous oleic acid (OA) into linoleic acid (LA). The disruption of the tauΔ12des in T. aureum by homologous recombination resulted in the accumulation of OA and a decrease in the levels of LA and its downstream PUFAs. However, the DHA content was increased slightly in tauΔ12des-disruption mutants, suggesting that DHA is primarily produced in T. aureum via the PUFA synthase pathway. The transformation of the tauΔ12des-disruption mutants with a tauΔ12des expression cassette restored the wild-type fatty acid profiles. These data clearly indicate that TauΔ12des functions as Δ12-fatty acid desaturase in the standard pathway of T. aureum and demonstrate that this thraustochytrid produces PUFAs via both the PUFA synthase and the standard pathways. |
topic |
polyunsaturated fatty acid polyketide synthase-like (PUFA synthase) pathway desaturase/elongase (standard) pathway homologous recombination targeted gene disruption |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520317144 |
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