A new peptide-n4-(acetyl-β-glucosaminyl)asparagine amidase from soybean (glycine max) seeds: Purification and substrate specificity

Yoshinobu Kimura, Akira Ohno

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We report here the isolation and characterization of a peptide-N4-(acetyl-β-glucosaminyl) asparagine amidase (peptide: N-glycanase) from soybean (Glycine max) seeds. The enzyme was purified to homogeneity with 6.5% yield from defatted soybean meal extract by ion-exchange chromatography, gel filtration, hydroxyapatite chromatography, and hydrophobic chromatography. The purified enzyme, designated PNGase-GM, had the apparent molecular mass of 93 kDa by SDS-PAGE and 90 kDa by gel filtration, indicating this PNGase is a monomeric protein. The enzyme showed maximal activity at pH 4.5-5.0. PNGase-GM was capable of hydrolyzing the β-aspartylglycosylamine linkage (GlcNAcβ1→Asn) of various glycopeptide substrates bearing high-mannose type, hybrid type, and xylose/fucose-containing plant complex type N-glycan units, while this amidase was far less active on the glycopeptides bearing sialylated animal complex-type gl.

Original languageEnglish
Pages (from-to)412-418
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume62
Issue number2
DOIs
Publication statusPublished - Jan 1 1998

Keywords

  • Free N-glycan
  • Glycine max
  • Glycoamidase
  • N-glycan releasing enzyme
  • Peptide: N-glycanase

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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