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Expression and Characterization of Human β-1, 4-Galactosyltransferase 1 (β4GalT1) Using Silkworm–Baculovirus Expression System

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Abstract

Baculovirus expression vector system (BEVS) is widely known as a mass-production tool to produce functional recombinant glycoproteins except that it may not be always suitable for medical practice due to the differences in the structure of N-linked glycans between insects and mammalian. Currently, various approaches have been reported to alter N-linked glycan structures of glycoproteins derived from insects into terminally sialylated complex-type N-glycans. In the light of those studies, we also proposed in vitro maturation of N-glycan with mass-produced and purified glycosyltransferases by silkworm–BEVS. β-1,4-Galactosyltransferase 1 (β4GalT1) is known as one of type II transmembrane enzymes that transfer galactose in a β-1, 4 linkage to accepter sugars, and a key enzyme for further sialylation of N-glycans. In this study, we developed a large-scale production of recombinant human β4GalT1 (rhβ4GalT1) with N- or C-terminal tags in silkworm–BEVS. We demonstrated that rhβ4GalT1 is N-glycosylated and without mucin-type glycosylation. Interestingly, we found that purified rhβ4GalT1 from silkworm serum presented higher galactosyltransferase activity than that expressed from cultured mammalian cells. We also validated the UDP-galactose transferase activity of produced rhβ4GalT1 proteins by using protein subtracts from silkworm silk gland. Taken together, rhβ4GalT1 from silkworms can become a valuable tool for producing high-quality recombinant glycoproteins with mammalian-like N-glycans.

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Authors and Affiliations

  1. Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki 6-10-1, Higashi-ku, Fukuoka, 812-8581, Japan

    Daisuke Morokuma, Jian Xu, Masato Hino, Hiroaki Mon, Takahiro Kusakabe & Jae Man Lee

  2. Laboratory of Cell Migration and Signaling, Division of Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology, 4700 KAUST Thuwal, Jeddah, 23955, Saudi Arabia

    Jasmeen S. Merzaban

  3. Laboratory of DNA Replication and Recombination, Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, 4700 KAUST Thuwal, Jeddah, 23955, Saudi Arabia

    Masateru Takahashi

Authors
  1. Daisuke Morokuma
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  2. Jian Xu
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  3. Masato Hino
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  4. Hiroaki Mon
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  5. Jasmeen S. Merzaban
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  6. Masateru Takahashi
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  7. Takahiro Kusakabe
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  8. Jae Man Lee
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Correspondence to Jae Man Lee.

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Morokuma, D., Xu, J., Hino, M. et al. Expression and Characterization of Human β-1, 4-Galactosyltransferase 1 (β4GalT1) Using Silkworm–Baculovirus Expression System. Mol Biotechnol 59, 151–158 (2017). https://doi.org/10.1007/s12033-017-0003-1

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  • DOI: https://doi.org/10.1007/s12033-017-0003-1

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