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Heterologous Production and Glycosylation of Japanese Eel Follitropin Using Silkworm

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  • Animal Cell Biotechnology
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Abstract

Follitropin, an important gonadotropin hormone, participates in vitellogenesis and spermatogenesis. Equine chorionic gonadotropin (eCG) can induce gonadotropin hormone activity in non-equid species and exhibits a long biological half-life. Here, we report the production, using silkworm larval and pupal systems, of biologically active recombinant hybrid-type follitropins based on the coding sequence of the eCG C-terminal peptide (CTP) between the mature β- and α-chains of eel. The three constructs, rJeFSH, rJeFSH·eCG, and rJeFSH·2xeCG were produced and verified to be N- or O-glycosylated and secreted mature peptides. Although rJeFSH·eCG contains more elaborate O-linked carbohydrate chains than rJeFSH, it elicited no significant in vitro oocyte maturation, which may be a result of insufficient terminal sialylation of its N-and O-linked carbohydrate chains. Then, a hybrid of rJeFSH·2xeCG extended with two eCG CTP. Furthermore, the receptor binding assay revealed potency of rJeFSH and rJeFSH·2xeCG to be a few folds greater than that of rJeFSH·eCG. The findings of this study will be useful for the development of more efficient GTHs in teleosts, including eels, when various modifications with two or more extended eCG CTP produced by silkworm are included.

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Acknowledgments

This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bio industry Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA, 11803422) and the National Institute of Fisheries Science (NIFS; R2019031).

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Authors and Affiliations

  1. Research and Development Department, Gyeongbuk Institute for Marine Bioindustry, Uljin, 36315, Korea

    Sun Mee Hong, Ji-Hyun Choi & Sun-Jung Jo

  2. Animal Biotechnology, Graduate School of Future Convergence Technology, Institute of Genetic Engineering, Hankyong National University, Anseong, 17579, Korea

    Kwan-Sik Min

  3. Jeju Fisheries Research Division, National Institute of Fisheries Science, Jeju, 63068, Korea

    Dae-Jung Kim

  4. Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan

    Jae Man Lee & Takahiro Kusakabe

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  1. Sun Mee Hong
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  2. Ji-Hyun Choi
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Hong, S.M., Choi, JH., Jo, SJ. et al. Heterologous Production and Glycosylation of Japanese Eel Follitropin Using Silkworm. Biotechnol Bioproc E 24, 745–753 (2019). https://doi.org/10.1007/s12257-019-0045-2

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  • DOI: https://doi.org/10.1007/s12257-019-0045-2

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