Peptide-N-glycosidase F Peptide-N4-（acetyl-β-glucosaminyl）-asparagine amidase
 

Contents

Solution in 50 mM sodium phosphate, 12.5 mM EDTA, 50% glycerol （v/v）, pH 7.2

Use N-glycosidase F to cleave all types of asparagine-bound N-glycans, provided that the amino group as well as the carboxyl group are present in a peptide linkage, and that the oligosaccharide has the minimum length of the chitobiose core unit. The reaction products are ammonia, aspartic acid （in the peptide chain）, and the complete oligosaccharide.
Note: N-Glycosidase F, recombinant is also available as a lyophilizate without glycerol.

EC 3.5.1.52
Optimum pH: 7.0 - 8.0
Molecular weight: M_r = 35.5 kD
Specific activity: 25,000 U/mg enzyme protein.
1 unit is the enzyme activity which hydrolyzes 1 nmol dabsyl fibrin glycopeptide or 0.2 nmol dansyl fetuin glycopeptide within 1 minute at +37°C and pH 7.8
Specificity: Hydrolyzes all types of N-glycan chains from glycopeptides and glycoproteins unless they carry α-1,3 linked core fucose residues present in insect and plant glycoproteins. Free of contaminating proteolytic activities [x = H or sugar（s）].
 

Principle

The reaction mechanism differs from that of endoglycosidases D, H, and F. These enzymes cleave the glycosidic linkage between the two N-acetylglucosamine residues. They also show a more limited substrate specificity than N-glycosidase F. Peptide-N-glycosidase A from Almond emulsin shows a similar substrate specificity as N-glycosidase F, but is often not able to efficiently remove all susceptible oligosaccharides.
 

Quality

Absence of contaminants: endoglycosidase F, β-galatctosidase, β-glucosidase, α- and β-mannosidase, β-N-acetylhexosaminidase, α-L-fucosidase, sialidase and proteases are not detectable.