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A novel membrane-anchored cytochrome c-550 of alkaliphilic Bacillus clarkii K24-1U: expression, molecular features and properties of redox potential

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Abstract

A membrane-anchored cytochrome c-550, which is highly expressed in obligately alkaliphilic Bacillus clarkii K24-1U, was purified and characterized. The protein contained a conspicuous sequence of Gly22-Asn34, in comparison with the other Bacillus small cytochromes c. Analytical data indicated that the original and lipase-treated intermediate forms of cytochrome c-550 bind to fatty acids of C15, C16 and C17 chain lengths and C15 chain length, respectively, and it was considered that these fatty acids are bound to glycerol–Cys18. Since there was a possibility that the presence of a diacylglycerol anchor contributed to the formation of dimeric states of this protein (20 and 17 kDa in SDS-PAGE), a C18M (Cys18 → Met)-cytochrome c-550 was constructed. The molecular mass of the C18M-cytochrome c-550 was determined as 15 and 10 kDa in SDS-PAGE and 23 kDa in blue native PAGE. The C18M-cytochrome c-550 bound with or without Triton X-100 formed a tetramer as the original cytochrome c-550 bound with Triton X-100, as determined by gel filtration. The midpoint redox potential of cytochrome c-550 as determined by redox titration was +83 mV, while that determined by cyclic voltammetric measurement was +7 mV. The above results indicate that cytochrome c-550 is a novel cytochrome c.

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Authors and Affiliations

  1. Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Tsukisamu-Higashi, Toyohira-ku, Sapporo, 062-8517, Japan

    Shinichi Ogami, Shoichi Hijikata, Tamotsu Tsukahara, Yasuhiro Mie, Toshihide Matsuno, Naoki Morita, Isao Hara, Tamotsu Hoshino, Kazuaki Yoshimune & Isao Yumoto

  2. Graduate School of Agriculture, Hokkaido University, Kita-ku, Sapporo, 060-8589, Japan

    Shinichi Ogami, Shoichi Hijikata, Toshihide Matsuno & Isao Yumoto

  3. Faculty of Fisheries, Hokkaido University, Hakodate, 041-0821, Japan

    Tamotsu Tsukahara, Koji Yamazaki & Norio Inoue

  4. Research Faculty of Agriculture, Hokkaido University, Kita-ku, Sapporo, 060-8589, Japan

    Atsushi Yokota

  5. Graduate School of Science, Hokkaido University, Kita-ku, Sapporo, 060-0810, Japan

    Tamotsu Hoshino

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  1. Shinichi Ogami
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Correspondence to Isao Yumoto.

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Communicated by A. Driessen.

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Ogami, S., Hijikata, S., Tsukahara, T. et al. A novel membrane-anchored cytochrome c-550 of alkaliphilic Bacillus clarkii K24-1U: expression, molecular features and properties of redox potential. Extremophiles 13, 491–504 (2009). https://doi.org/10.1007/s00792-009-0234-6

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