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Lack of ∆5 Desaturase Activity Impairs EPA and DHA Synthesis in Fish Cells from Red Sea Bream and Japanese Flounder

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Abstract

Long-chain (≥ C20) polyunsaturated fatty acids (LC-PUFA), such as eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA), are necessary for human health and are obtained from marine fish-derived oils. Marine fish are LC-PUFA-rich animals; however, many of them require LC-PUFA for growth. Therefore, it is suggested that they do not have sufficient ability to biosynthesize LC-PUFA. To evaluate in vivo LC-PUFA synthetic activity in fish cells, fish-derived cell lines from red sea bream (Pagrus major, PMS and PMF), Japanese flounder (Paralichthys olivaceus, HINAE), and zebrafish (Danio rerio, BRF41) were incubated with n-3 fatty acids labeled by radioisotopes or stable isotopes, and then, n-3 PUFA were analyzed by thin-layer chromatography or liquid chromatography-mass spectrometry. Labeled EPA and DHA were biosynthesized from labeled α-linolenic acid (18:3n-3) in BRF41, whereas they were not detected in PMS, PMF, or HINAE cells. We next cloned the fatty acid desaturase 2 (Fads2) cDNAs from PMF cells and zebrafish, expressed in budding yeasts, and then analyzed the substrate specificities of enzymes. As a result, we found that Fads2 from PMF cells was a ∆6/∆8 desaturase. Collectively, our study indicates that cell lines from red sea bream and Japanese flounder were not able to synthesize EPA or DHA by themselves, possibly due to the lack of ∆5 desaturase activity. Furthermore, this study provides a sensitive and reproducible non-radioactive method for evaluating LC-PUFA synthesis in fish cells using a stable isotope and liquid chromatography-mass spectrometry.

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Availability of Data and Material

All data not present in the manuscript is available upon request (correspondence to N.O., nokino@agr.kyushu-u.ac.jp).

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Acknowledgements

We thank Dr. Jun Kurita (National Research Institute of Aquaculture, Fisheries Research and Education Agency) for providing us the PMS and PMF cell lines. We are also grateful to Dr. Hisae Kasai (Faculty of Fisheries Sciences, Hokkaido University) who provided us the HINAE cell line.

Funding

This work was supported in part by the Sasakawa Scientific Research Grant from The Japan Science Society (to H.Y.).

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 744 Moto-oka, Nishi-ku, Fukuoka, 819-0395, Japan

    Hayato Nyunoya, Tatsuki Noda, You Kawamoto, Yasuhiro Hayashi, Yohei Ishibashi, Makoto Ito & Nozomu Okino

  2. Faculty of Pharma-Science, Teikyo University, Kaga 2-11-1, Itabashi-ku, Tokyo, 173-8605, Japan

    Yasuhiro Hayashi

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  1. Hayato Nyunoya
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  2. Tatsuki Noda
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Correspondence to Nozomu Okino.

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Nyunoya, H., Noda, T., Kawamoto, Y. et al. Lack of ∆5 Desaturase Activity Impairs EPA and DHA Synthesis in Fish Cells from Red Sea Bream and Japanese Flounder. Mar Biotechnol 23, 472–481 (2021). https://doi.org/10.1007/s10126-021-10040-9

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