Expression, purification, and characterization of endo-β-N- acetylglucosaminidase H using baculovirus-mediated silkworm protein expression system

Takumi Mitsudome, Jian Xu, Yudai Nagata, Atsushi Masuda, Kazuhiro Iiyama, Daisuke Morokuma, Zhiqing Li, Hiroaki Mon, Jae Man Lee, Takahiro Kusakabe

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

Endo-β-N-acetylglucosaminidase (Endo H) from Streptomyces plicatus hydrolyzes the core di-GlcNAc units of asparagine-linked oligosaccharides and is regarded as an important tool for glycobiology research. In the present study, we established a large-scale system to produce secreted Endo H using a silkworm-baculovirus expression system (silkworm-BES). The recombinant Endo H purified from silkworm hemolymph had activity comparable to that from recombinant Escherichia coli. As well as its well-characterized substrate RNase B, the Endo H from silkworm-BES was able to deglycosylate the high-mannose glycoproteins from silkworm hemolymph. Interestingly, the secretion amount of recombinant Endo H was significantly varied among the different silkworm strains, which could provide valuable information for larger-scale protein productions from silkworm-BES.

Original languageEnglish
Pages (from-to)3978-3988
Number of pages11
JournalApplied Biochemistry and Biotechnology
Volume172
Issue number8
DOIs
Publication statusPublished - Apr 2014

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

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