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KMID : 1094720160210030381
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 3 p.381 ~ p.388
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Molecular analysis and bioactivity of luteinizing hormone from Japanese eel, Anguilla japonica, produced in silkworm pupae
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Choi Ji-Hyun
Kim Dae-Jung
Hong Sun-Mee
Jo Sun-Jung
Min Kwan-Sik
Sohn Young-Chang
Lee Jae-Man
Kusakabe Takahiro
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Abstract
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Luteinizing hormone (LH), a gonadotropin hormone (GTH) of the pituitary glycoprotein family, is important in oocyte maturation, ovulation, and spermiation. In this study, we generated a Japanese eel LH (JeLH) with and without the equine chorionic gonadotropin (eCG) carboxyl-terminal peptides under the control of the polyhedron in the silkworm pupae BES to determine their expression levels, glycosylation profile, and in vitro bioactivity. The target proteins were highly expressed in the pupae hemolymph. Recombinant JeLH·eCG and JeLH were N- or O-glycosylated, as shown by periodic-acid Schiff staining, deglycosidase enzyme treatment, and lectin blot analyses, and showed no significant difference in the in vitro bioactivity. Both single-chain hormones and salmon pituitary extract induced maturation of Japanese eel oocytes in vitro. Recombinant LHs produced in silkworm pupae might be suitable candidates for in vivo experiments, because they can be produced in sufficient amount and can undergo N-glycosylation.
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KEYWORD
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Japanese eel, luteinizing hormone, silkworm, recombinant protein, equine chorionic gonadotropin
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