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Possible Biosynthetic Pathways for all cis-3,6,9,12,15,19,22,25,28-Hentriacontanonaene in Bacteria

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Lipids

Abstract

A very long chain polyunsaturated hydrocarbon, hentriacontanonaene (C31:9), was detected in an eicosapentaenoic acid (EPA)-producing marine bacterium, which was isolated from the mid-latitude seashore of Hokkaido, Japan, and was tentatively identified as mesophilic Shewanella sp. strain osh08 from 16S rRNA gene sequencing. The geometry and position of the double bonds in this compound were determined physicochemically to be all cis at positions 3, 6, 9, 12, 15, 19, 22, 25, and 28. Although C31:9 was detected in all of the seven EPA- or/and docosahexaenoic acid-producing bacteria tested, an EPA-deficient mutant (strain IK-1Δ8) of one of these bacteria had no C31:9. Strain IK-1Δ8 had defects in the pfaD gene, one of the five pfa genes responsible for the biosynthesis of EPA. Although Escherichia coli DH5α does not produce EPA or DHA inherently, cells transformed with the pfa genes responsible for the biosynthesis of EPA and DHA produced EPA and DHA, respectively, but not C31:9. These results suggest that the Pfa protein complex is involved in the biosynthesis of C31:9 and that pfa genes must not be the only genes responsible for the formation of C31:9. In this report, we determined for the first time the molecular structure of the C31:9 and discuss the possible biosynthetic pathways of this compound.

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Abbreviations

CI:

Chemical ionization

CI-GC/MS:

Chemical ionization–mass spectrometry

DHA:

Docosahexaenoic acid

EI:

Electron impact ionization

EI-GC/MS:

Electron impact ionization–mass spectrometry

EPA:

Eicosapentaenoic acid

FAME:

Fatty acid methyl ester

GC/MS:

Gas chromatography–mass spectrometry

GLC:

Gas–liquid chromatography

HPLC:

High-performance liquid chromatography

PUHC:

Polyunsaturated hydrocarbon

PUFA:

Polyunsaturated fatty acid

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Acknowledgments

The infrared analysis of C31:9 was carried out using the apparatus of the OPEN FACILITY, Hokkaido University Sousei Hall with extensive assistance by and discussion with Dr. Y. Matsuo. The authors would like to express their appreciation to Drs. K. Shimazu and K. Nakata of the Faculty of Environmental Earth Science, Hokkaido University for their discussions throughout the work. This work was partly supported by a grant from the National Institute of Polar Research and a Grant-in-Aid from Hokkaido Innovation through Nanotechnology Support (HINTS).

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Author notes

  1. Kazuo Watanabe

    Present address: WysiWyg Co., Ltd., Haccobori, Chuo-ku, Tokyo, 104-0032, Japan

Authors and Affiliations

  1. Course in Environmental Molecular Biology and Microbial Ecology, Division of Biosphere Science, Graduate School of Environmental Science, Hokkaido University, Kita-ku, Sapporo, 060-0810, Japan

    Shinji Sugihara, Hitomi Nakanowatari & Hidetoshi Okuyama

  2. Department of Biological Sciences, Faculty of Science, Hokkaido University, Kita-ku, Sapporo, 060-0810, Japan

    Ryuji Hori & Hidetoshi Okuyama

  3. Laboratory of Environmental Molecular Biology, Faculty of Environmental Earth Science, Hokkaido University, Kitaku-ku, Sapporo, 060-0810, Japan

    Ryuji Hori & Hidetoshi Okuyama

  4. Division of Cellular Life Science, Faculty of Advanced Life Science, Hokkaido University, Kita-ku, Sapporo, 060-0810, Japan

    Yasuhiro Takada

  5. Research Institute of Genome-Based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Toyohira-ku, Sapporo, 062-8517, Japan

    Isao Yumoto & Naoki Morita

  6. National Research Institute of Fisheries Science, Fisheries Research Agency, Kanazawa-ku, Yokohama, 236-8648, Japan

    Yutaka Yano

  7. Microbial Engineering Group, Sagami Chemical Research Center, Hayakawa, Ayase, 252-1193, Japan

    Kazuo Watanabe

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  1. Shinji Sugihara
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  9. Hidetoshi Okuyama
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Correspondence to Hidetoshi Okuyama.

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Sugihara, S., Hori, R., Nakanowatari, H. et al. Possible Biosynthetic Pathways for all cis-3,6,9,12,15,19,22,25,28-Hentriacontanonaene in Bacteria. Lipids 45, 167–177 (2010). https://doi.org/10.1007/s11745-009-3380-9

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