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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Main Authors: Takanori Matsuda, Keishi Sakaguchi, Rie Hamaguchi, Takumi Kobayashi, Eriko Abe, Yoichiro Hama, Masahiro Hayashi, Daiske Honda, Yuji Okita, Shinichi Sugimoto, Nozomu Okino, Makoto Ito
Format: Article
Language:English
Published: Elsevier 2012-06-01
Series:Journal of Lipid Research
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S0022227520317144
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spelling 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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