PMID: 9532807Apr 9, 1998Paper

A new peptide-N4-(acetyl-beta-glucosaminyl)asparagine amidase from soybean (Glycine max) seeds: purification and substrate specificity

Bioscience, Biotechnology, and Biochemistry
Y Kimura, A Ohno

Abstract

We report here the isolation and characterization of a peptide-N4-(acetyl-beta-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 beta-aspartylglycosylamine linkage (GlcNAc beta 1-->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 glycans.

References

Dec 1, 1978·Journal of Biochemistry·N Takahashi, H Nishibe
Feb 16, 1987·European Journal of Biochemistry·T H PlummerA L Tarentino
Jun 1, 1988·Journal of Biochemistry·Y KimuraG Funatsu
Apr 15, 1983·Biochemical and Biophysical Research Communications·K SugiyamaN Takahashi
Dec 24, 1997·Bioscience, Biotechnology, and Biochemistry·Y KimuraS Takagi
Jan 1, 1997·Bioscience, Biotechnology, and Biochemistry·Yoshinobu KimuraTomonori Shiraishi

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Citations

Nov 11, 2008·Bioscience, Biotechnology, and Biochemistry·Kosuke NakamuraYoshinobu Kimura
Aug 10, 2006·Glycoconjugate Journal·Tadashi Suzuki, Yoko Funakoshi
Aug 19, 2009·Biomedical Chromatography : BMC·Sze Kwan Lam, Tzi Bun Ng
Mar 17, 2015·Cellular and Molecular Life Sciences : CMLS·Yoichiro HaradaTadashi Suzuki
Apr 17, 2018·Bioscience, Biotechnology, and Biochemistry·Ryota UemuraYoshinobu Kimura
Dec 12, 2017·Mass Spectrometry Reviews·Qiwei ZhangJianjun Li
Oct 31, 2000·Bioscience, Biotechnology, and Biochemistry·Y Kimura, E Kitahara
Jun 22, 2006·Journal of Biochemistry·Takeshi IshimizuSumihiro Hase
Dec 25, 2004·Bioscience, Biotechnology, and Biochemistry·Kwan Kit WooYoshinobu Kimura
Apr 19, 2002·Biochimica Et Biophysica Acta·Yoshinobu KimuraMikio Nishimura

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