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Isolation and purification of the major photosynthetic antenna, fucoxanthin-Chl a/c protein, from cultured discoid germilings of the brown Alga, Cladosiphon okamuranus TOKIDA (Okinawa Mozuku)

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Abstract

A chlorophyll c binding membrane intrinsic light-harvesting complex, the fucoxanthin-chlorophyll a/c protein (FCP), was isolated from cultured discoid germilings of an edible Japanese brown alga, Cladosiphon (C.) okamuranus TOKIDA (Okinawa Mozuku in Japanese). The discoid germiling is an ideal source of brown algal photosynthetic pigment-protein complexes in terms of its size and easiness of cultivation on a large scale. Ion-exchange chromatography was crucial for the purification of FCP from solubilized thylakoid proteins. The molecular weight of the purified FCP assembly was estimated to be ~56 kDa using blue native-PAGE. Further subunit analyses using 2D-PAGE revealed that the FCP assembled as a trimer consisting of two distinguishable subunits having molecular weights of 18.2 (H) and 17.5 (L) kDa. Fluorescence and fluorescence-excitation spectra confirmed that the purified FCP assembly was functionally intact.

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Abbreviations

BN-PAGE:

Blue native-PAGE

C. :

Cladosiphon

CBB:

Coomassie brilliant blue

Chl:

Chlorophyll

DM:

n-dodecyl-β-d-maltopyranoside

FCP:

The fucoxanthin-chlorophyll a/c protein

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Acknowledgments

RF and MK thank Dr. Yasufumi Umena (Osaka City Univ.), Dr. Keisuke Kawakami (Osaka City Univ.) and Prof. Dr. Jian-Ren Shen (Okayama Univ.) for kind introduction and discussion of blue native (BN)-PAGE. RF and MK thank Prof. Dr. Makoto Miyata (Osaka City Univ.) for kind support to prepare the sample for N-terminal sequence. This work is supported by a grant-in-aid from Japanese Science and Technology Agency (JST), A-step Feasibility Study # AS2111259E.

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Authors and Affiliations

  1. The OCU Advanced Research Institute for Natural Science and Technology (OCARINA), 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka, 558-8585, Japan

    Ritsuko Fujii & Hideki Hashimoto

  2. CREST/JST, 4-1-8 Hon-cho Kawaguchi, Saitama, 332-0012, Japan

    Ritsuko Fujii, Mamiko Kita & Hideki Hashimoto

  3. Department of Physics, Graduate School of Science, Osaka City University, 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka, 558-8585, Japan

    Mamiko Kita & Hideki Hashimoto

  4. South Product Co. Ltd, 12-75 Suzaki, Uruma, Okinawa, 904-2234, Japan

    Yoshiro Iinuma, Naohiro Oka, Yuki Takaesu, Tomonori Taira & Masahiko Iha

  5. Glasgow Biomedical Research Centre, University of Glasgow, 12 University Place, Glasgow, G12 8QQ, Scotland, UK

    Richard J. Cogdell

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  1. Ritsuko Fujii
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  2. Mamiko Kita
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Correspondence to Ritsuko Fujii or Hideki Hashimoto.

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Fujii, R., Kita, M., Iinuma, Y. et al. Isolation and purification of the major photosynthetic antenna, fucoxanthin-Chl a/c protein, from cultured discoid germilings of the brown Alga, Cladosiphon okamuranus TOKIDA (Okinawa Mozuku). Photosynth Res 111, 157–163 (2012). https://doi.org/10.1007/s11120-011-9688-3

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