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Expression and characterization of rainbow trout Oncorhynchus mykiss recombinant myoglobin

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Abstract

Recombinant expression system was established for rainbow trout myoglobin (Mb) considering its unique primary structure of having one unusual deletion and two cysteine residues in contrast to the other fish Mbs. The obtained recombinant Mb without His-tag showed non-cooperative thermal denaturation profile. The presence of free cysteine residue(s) in rainbow trout Mb was demonstrated by reacting with a sulfhydryl agent, 4, 4´-dithiodipyridine, which ultimately resulted in the oxidation of Mb with characteristic changes in visible absorption spectra. Besides, the recombinant Mb displayed steady peroxidase reactivity indicating in vivo roles of Mb as a reactive oxygen species scavenger. The findings of the present study indicate that the solitary rainbow trout Mb, which ultimately manifest typical secondary structure pattern and corroborate characteristic functionality, can be over expressed in recombinant system devoid of fusion tag.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was partly supported by the Japan Society for Promotion of Sciences (KAKENHI # 22380015 to Y. O.).

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Authors and Affiliations

  1. Laboratory of Marine Biochemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, 113-8657, Japan

    Muhammad Mehedi Hasan & Hideki Ushio

  2. Department of Fisheries Technology, Bangladesh Agricultural University, Mymensingh, 2202, Bangladesh

    Muhammad Mehedi Hasan

  3. Graduate School of Agriculture, Tohoku University, Aramaki, Aoba, Sendai, 980-8572, Japan

    Yoshihiro Ochiai

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  1. Muhammad Mehedi Hasan
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  2. Hideki Ushio
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  3. Yoshihiro Ochiai
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All authors planned and designed the studies. Material preparation, data collection, and analyses were performed by MH. MH wrote the manuscript and all authors read and approved the submitted version.

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Correspondence to Muhammad Mehedi Hasan.

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Hasan, M.M., Ushio, H. & Ochiai, Y. Expression and characterization of rainbow trout Oncorhynchus mykiss recombinant myoglobin. Fish Physiol Biochem 47, 1477–1488 (2021). https://doi.org/10.1007/s10695-021-00991-0

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